INTRODUCTION TO PHARMACOLOGY

Pharmacology is the study of how chemicals

interact with the body

Endogenous ndash hormones growth factors etc

Exogenous ndash drugs

Two areas of study

Pharmacodynamics

Interaction of chemicals with receptors or enzymes

Pharmacokinetics

Absorption distribution metabolism excretion

1

Pharmacokinetics

ldquoWhat the body does to a drugrdquo

LADME

Liberation

Absorption

Distribution

Metabolism

Excretion

2

Pharmacokinetics

Routes of drug administration

Oral

Sublingual

Intravenous (IV)

Intraperitoneal (IP)

Intramuscular (IM)

Subcutaneous (SC)

Rectal

Epidural

Intracerebroventricular

3

Pharmacokinetics

Liberation

How is a drug released from its delivery vehicle

Pills must dissolve or be broken down

Forms of oral medications

Tablet

Caplet

Liquid

Liqui-Gel

4

Pharmacokinetics

Absorption

How is a drug absorbed into the tissue or

bloodstream

Depends on route of administration

Oral ndash absorbed via gastrointestinal tract goes to liver

Sublingual ndash absorbed via blood vessels under the

tongue bypasses the liver

IV ndash na

IP ndash absorbed via peritoneal mucosae

IM ndash absorbed via muscle lymphatics amp capillaries

Bioavailability how much ingested drug is

actually absorbed

5

For an orally-administered medication

absorption can only occur once which of these

other processes has occurred

A) Distribution

B) Metabolism

C) Liberation

D) Excretion

6

Why does the previous question include the

qualifer ldquoFor an orally-administered

medicationrdquo

7

Pharmacokinetics

Distribution

How is a drug distributed to the various fluid

compartments or tissues of the body

Plasma vs interstitial vs intracellular fluid

Adipose tissue vs lean tissues

Distribution is unequal amp distinct for each drug

ldquoVolume of distributionrdquo ndash theoretical volume a drug

would occupy if it were present in all compartments at

the same concentration as in the plasma

8

ldquoVolume of distributionrdquo is based MOSTLY onhellip

A) how much blood a patient has

B) how much drug was in a particular pill

C) how soluble a particular drug is in lipids

vs water

9

True or false The volume of distribution of a

drug may be larger than the total volume of the

patient

A) True

B) False

10

Pharmacokinetics

Metabolism

Is a drug chemically modified by the body If sohellip

Are the metabolites more or less active

Are the metabolites safer or more toxic

Can the metabolites be more easily excreted

Which tissues metabolize the drug

What enzyme systems or other reactions are used to

metabolize the drug

11

Pharmacokinetics

Metabolism

Phase I reactions

Liver

Cytochrome P450 (cyp450) enzymes

Eg Oxidation reduction hydrolysis (de)cyclization

Often produce active metabolites

First-pass effect

Phase II reactions

Liver and kidney

Addition of polar functional groups

Usually produce non-functional metabolites for excretion

Eg Glucuronidation glutathione conjugation

12

Pharmacokinetics

Metabolic clearance of drugs hormones

Metabolized into non-active form

Bound to cells amp degraded recycled

Excreted

These processes are not mutually exclusive

Drugs can be metabolized to inactive metabolites then

excreted

13

Pharmacokinetics

Metabolic clearance of drugs hormones

Rate of removal depends on solubility

Peptide hormones

Degraded in blood or tissues

Cleared rapidly (seconds to minutes)

Steroid hormones

Bounds to plasma proteins

Cleared slowly (minutes to days)

14

Drug Hormone Clearance

Metabolic clearance rate (MCR)

Rate of disappearance from plasma

Infuse at increasing rate until plasma concentration

reaches steady-state

Infusion rate = rate of disappearance

Concentration of hormone in plasma

15

MCR (mLmin) = rate of hormone disappearance (ngmin)

Concentration of hormone (ngmL)

Pharmacokinetics

Excretion Elimination

Urine

Feces

Breath

Skin

16

Metabolic clearance rate refers tohellip

A) How fast kidneys produce urine

B) How much drug is filtered by the kidneys

C) A theoretical volume of plasma that can be

100 cleared of drug in a particular amount of

time

17

Pharmacodynamics

ldquoWhat a drug does to the bodyrdquo

Drug (ligand)-receptor interactions

Receptor types ndash membrane vs intracellular

Enzymes amp structural proteins can act as drug ldquoreceptorsrdquo

Signal transduction pathways

Enzyme-based biochemical pathways

18

L + R L R

Pharmacodynamics

Potential actionsroles of drugs

Agonist

Stimulating action vs depressing action

Full or partial agonist

Antagonist

Direct beneficial chemical reaction

Direct harmful chemical reaction

19

Pharmacodynamics

Dose-response curves

Increased [drug] increased effect

EC50 (effective dose yielding 50 max effect)

Curve shifts

Upward ndash higher efficacy

Leftward ndash higher potency (sensitivity)

20

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Pharmacokinetics

ldquoWhat the body does to a drugrdquo

LADME

Liberation

Absorption

Distribution

Metabolism

Excretion

2

Pharmacokinetics

Routes of drug administration

Oral

Sublingual

Intravenous (IV)

Intraperitoneal (IP)

Intramuscular (IM)

Subcutaneous (SC)

Rectal

Epidural

Intracerebroventricular

3

Pharmacokinetics

Liberation

How is a drug released from its delivery vehicle

Pills must dissolve or be broken down

Forms of oral medications

Tablet

Caplet

Liquid

Liqui-Gel

4

Pharmacokinetics

Absorption

How is a drug absorbed into the tissue or

bloodstream

Depends on route of administration

Oral ndash absorbed via gastrointestinal tract goes to liver

Sublingual ndash absorbed via blood vessels under the

tongue bypasses the liver

IV ndash na

IP ndash absorbed via peritoneal mucosae

IM ndash absorbed via muscle lymphatics amp capillaries

Bioavailability how much ingested drug is

actually absorbed

5

For an orally-administered medication

absorption can only occur once which of these

other processes has occurred

A) Distribution

B) Metabolism

C) Liberation

D) Excretion

6

Why does the previous question include the

qualifer ldquoFor an orally-administered

medicationrdquo

7

Pharmacokinetics

Distribution

How is a drug distributed to the various fluid

compartments or tissues of the body

Plasma vs interstitial vs intracellular fluid

Adipose tissue vs lean tissues

Distribution is unequal amp distinct for each drug

ldquoVolume of distributionrdquo ndash theoretical volume a drug

would occupy if it were present in all compartments at

the same concentration as in the plasma

8

ldquoVolume of distributionrdquo is based MOSTLY onhellip

A) how much blood a patient has

B) how much drug was in a particular pill

C) how soluble a particular drug is in lipids

vs water

9

True or false The volume of distribution of a

drug may be larger than the total volume of the

patient

A) True

B) False

10

Pharmacokinetics

Metabolism

Is a drug chemically modified by the body If sohellip

Are the metabolites more or less active

Are the metabolites safer or more toxic

Can the metabolites be more easily excreted

Which tissues metabolize the drug

What enzyme systems or other reactions are used to

metabolize the drug

11

Pharmacokinetics

Metabolism

Phase I reactions

Liver

Cytochrome P450 (cyp450) enzymes

Eg Oxidation reduction hydrolysis (de)cyclization

Often produce active metabolites

First-pass effect

Phase II reactions

Liver and kidney

Addition of polar functional groups

Usually produce non-functional metabolites for excretion

Eg Glucuronidation glutathione conjugation

12

Pharmacokinetics

Metabolic clearance of drugs hormones

Metabolized into non-active form

Bound to cells amp degraded recycled

Excreted

These processes are not mutually exclusive

Drugs can be metabolized to inactive metabolites then

excreted

13

Pharmacokinetics

Metabolic clearance of drugs hormones

Rate of removal depends on solubility

Peptide hormones

Degraded in blood or tissues

Cleared rapidly (seconds to minutes)

Steroid hormones

Bounds to plasma proteins

Cleared slowly (minutes to days)

14

Drug Hormone Clearance

Metabolic clearance rate (MCR)

Rate of disappearance from plasma

Infuse at increasing rate until plasma concentration

reaches steady-state

Infusion rate = rate of disappearance

Concentration of hormone in plasma

15

MCR (mLmin) = rate of hormone disappearance (ngmin)

Concentration of hormone (ngmL)

Pharmacokinetics

Excretion Elimination

Urine

Feces

Breath

Skin

16

Metabolic clearance rate refers tohellip

A) How fast kidneys produce urine

B) How much drug is filtered by the kidneys

C) A theoretical volume of plasma that can be

100 cleared of drug in a particular amount of

time

17

Pharmacodynamics

ldquoWhat a drug does to the bodyrdquo

Drug (ligand)-receptor interactions

Receptor types ndash membrane vs intracellular

Enzymes amp structural proteins can act as drug ldquoreceptorsrdquo

Signal transduction pathways

Enzyme-based biochemical pathways

18

L + R L R

Pharmacodynamics

Potential actionsroles of drugs

Agonist

Stimulating action vs depressing action

Full or partial agonist

Antagonist

Direct beneficial chemical reaction

Direct harmful chemical reaction

19

Pharmacodynamics

Dose-response curves

Increased [drug] increased effect

EC50 (effective dose yielding 50 max effect)

Curve shifts

Upward ndash higher efficacy

Leftward ndash higher potency (sensitivity)

20

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Pharmacokinetics

Routes of drug administration

Oral

Sublingual

Intravenous (IV)

Intraperitoneal (IP)

Intramuscular (IM)

Subcutaneous (SC)

Rectal

Epidural

Intracerebroventricular

3

Pharmacokinetics

Liberation

How is a drug released from its delivery vehicle

Pills must dissolve or be broken down

Forms of oral medications

Tablet

Caplet

Liquid

Liqui-Gel

4

Pharmacokinetics

Absorption

How is a drug absorbed into the tissue or

bloodstream

Depends on route of administration

Oral ndash absorbed via gastrointestinal tract goes to liver

Sublingual ndash absorbed via blood vessels under the

tongue bypasses the liver

IV ndash na

IP ndash absorbed via peritoneal mucosae

IM ndash absorbed via muscle lymphatics amp capillaries

Bioavailability how much ingested drug is

actually absorbed

5

For an orally-administered medication

absorption can only occur once which of these

other processes has occurred

A) Distribution

B) Metabolism

C) Liberation

D) Excretion

6

Why does the previous question include the

qualifer ldquoFor an orally-administered

medicationrdquo

7

Pharmacokinetics

Distribution

How is a drug distributed to the various fluid

compartments or tissues of the body

Plasma vs interstitial vs intracellular fluid

Adipose tissue vs lean tissues

Distribution is unequal amp distinct for each drug

ldquoVolume of distributionrdquo ndash theoretical volume a drug

would occupy if it were present in all compartments at

the same concentration as in the plasma

8

ldquoVolume of distributionrdquo is based MOSTLY onhellip

A) how much blood a patient has

B) how much drug was in a particular pill

C) how soluble a particular drug is in lipids

vs water

9

True or false The volume of distribution of a

drug may be larger than the total volume of the

patient

A) True

B) False

10

Pharmacokinetics

Metabolism

Is a drug chemically modified by the body If sohellip

Are the metabolites more or less active

Are the metabolites safer or more toxic

Can the metabolites be more easily excreted

Which tissues metabolize the drug

What enzyme systems or other reactions are used to

metabolize the drug

11

Pharmacokinetics

Metabolism

Phase I reactions

Liver

Cytochrome P450 (cyp450) enzymes

Eg Oxidation reduction hydrolysis (de)cyclization

Often produce active metabolites

First-pass effect

Phase II reactions

Liver and kidney

Addition of polar functional groups

Usually produce non-functional metabolites for excretion

Eg Glucuronidation glutathione conjugation

12

Pharmacokinetics

Metabolic clearance of drugs hormones

Metabolized into non-active form

Bound to cells amp degraded recycled

Excreted

These processes are not mutually exclusive

Drugs can be metabolized to inactive metabolites then

excreted

13

Pharmacokinetics

Metabolic clearance of drugs hormones

Rate of removal depends on solubility

Peptide hormones

Degraded in blood or tissues

Cleared rapidly (seconds to minutes)

Steroid hormones

Bounds to plasma proteins

Cleared slowly (minutes to days)

14

Drug Hormone Clearance

Metabolic clearance rate (MCR)

Rate of disappearance from plasma

Infuse at increasing rate until plasma concentration

reaches steady-state

Infusion rate = rate of disappearance

Concentration of hormone in plasma

15

MCR (mLmin) = rate of hormone disappearance (ngmin)

Concentration of hormone (ngmL)

Pharmacokinetics

Excretion Elimination

Urine

Feces

Breath

Skin

16

Metabolic clearance rate refers tohellip

A) How fast kidneys produce urine

B) How much drug is filtered by the kidneys

C) A theoretical volume of plasma that can be

100 cleared of drug in a particular amount of

time

17

Pharmacodynamics

ldquoWhat a drug does to the bodyrdquo

Drug (ligand)-receptor interactions

Receptor types ndash membrane vs intracellular

Enzymes amp structural proteins can act as drug ldquoreceptorsrdquo

Signal transduction pathways

Enzyme-based biochemical pathways

18

L + R L R

Pharmacodynamics

Potential actionsroles of drugs

Agonist

Stimulating action vs depressing action

Full or partial agonist

Antagonist

Direct beneficial chemical reaction

Direct harmful chemical reaction

19

Pharmacodynamics

Dose-response curves

Increased [drug] increased effect

EC50 (effective dose yielding 50 max effect)

Curve shifts

Upward ndash higher efficacy

Leftward ndash higher potency (sensitivity)

20

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Pharmacokinetics

Liberation

How is a drug released from its delivery vehicle

Pills must dissolve or be broken down

Forms of oral medications

Tablet

Caplet

Liquid

Liqui-Gel

4

Pharmacokinetics

Absorption

How is a drug absorbed into the tissue or

bloodstream

Depends on route of administration

Oral ndash absorbed via gastrointestinal tract goes to liver

Sublingual ndash absorbed via blood vessels under the

tongue bypasses the liver

IV ndash na

IP ndash absorbed via peritoneal mucosae

IM ndash absorbed via muscle lymphatics amp capillaries

Bioavailability how much ingested drug is

actually absorbed

5

For an orally-administered medication

absorption can only occur once which of these

other processes has occurred

A) Distribution

B) Metabolism

C) Liberation

D) Excretion

6

Why does the previous question include the

qualifer ldquoFor an orally-administered

medicationrdquo

7

Pharmacokinetics

Distribution

How is a drug distributed to the various fluid

compartments or tissues of the body

Plasma vs interstitial vs intracellular fluid

Adipose tissue vs lean tissues

Distribution is unequal amp distinct for each drug

ldquoVolume of distributionrdquo ndash theoretical volume a drug

would occupy if it were present in all compartments at

the same concentration as in the plasma

8

ldquoVolume of distributionrdquo is based MOSTLY onhellip

A) how much blood a patient has

B) how much drug was in a particular pill

C) how soluble a particular drug is in lipids

vs water

9

True or false The volume of distribution of a

drug may be larger than the total volume of the

patient

A) True

B) False

10

Pharmacokinetics

Metabolism

Is a drug chemically modified by the body If sohellip

Are the metabolites more or less active

Are the metabolites safer or more toxic

Can the metabolites be more easily excreted

Which tissues metabolize the drug

What enzyme systems or other reactions are used to

metabolize the drug

11

Pharmacokinetics

Metabolism

Phase I reactions

Liver

Cytochrome P450 (cyp450) enzymes

Eg Oxidation reduction hydrolysis (de)cyclization

Often produce active metabolites

First-pass effect

Phase II reactions

Liver and kidney

Addition of polar functional groups

Usually produce non-functional metabolites for excretion

Eg Glucuronidation glutathione conjugation

12

Pharmacokinetics

Metabolic clearance of drugs hormones

Metabolized into non-active form

Bound to cells amp degraded recycled

Excreted

These processes are not mutually exclusive

Drugs can be metabolized to inactive metabolites then

excreted

13

Pharmacokinetics

Metabolic clearance of drugs hormones

Rate of removal depends on solubility

Peptide hormones

Degraded in blood or tissues

Cleared rapidly (seconds to minutes)

Steroid hormones

Bounds to plasma proteins

Cleared slowly (minutes to days)

14

Drug Hormone Clearance

Metabolic clearance rate (MCR)

Rate of disappearance from plasma

Infuse at increasing rate until plasma concentration

reaches steady-state

Infusion rate = rate of disappearance

Concentration of hormone in plasma

15

MCR (mLmin) = rate of hormone disappearance (ngmin)

Concentration of hormone (ngmL)

Pharmacokinetics

Excretion Elimination

Urine

Feces

Breath

Skin

16

Metabolic clearance rate refers tohellip

A) How fast kidneys produce urine

B) How much drug is filtered by the kidneys

C) A theoretical volume of plasma that can be

100 cleared of drug in a particular amount of

time

17

Pharmacodynamics

ldquoWhat a drug does to the bodyrdquo

Drug (ligand)-receptor interactions

Receptor types ndash membrane vs intracellular

Enzymes amp structural proteins can act as drug ldquoreceptorsrdquo

Signal transduction pathways

Enzyme-based biochemical pathways

18

L + R L R

Pharmacodynamics

Potential actionsroles of drugs

Agonist

Stimulating action vs depressing action

Full or partial agonist

Antagonist

Direct beneficial chemical reaction

Direct harmful chemical reaction

19

Pharmacodynamics

Dose-response curves

Increased [drug] increased effect

EC50 (effective dose yielding 50 max effect)

Curve shifts

Upward ndash higher efficacy

Leftward ndash higher potency (sensitivity)

20

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Pharmacokinetics

Absorption

How is a drug absorbed into the tissue or

bloodstream

Depends on route of administration

Oral ndash absorbed via gastrointestinal tract goes to liver

Sublingual ndash absorbed via blood vessels under the

tongue bypasses the liver

IV ndash na

IP ndash absorbed via peritoneal mucosae

IM ndash absorbed via muscle lymphatics amp capillaries

Bioavailability how much ingested drug is

actually absorbed

5

For an orally-administered medication

absorption can only occur once which of these

other processes has occurred

A) Distribution

B) Metabolism

C) Liberation

D) Excretion

6

Why does the previous question include the

qualifer ldquoFor an orally-administered

medicationrdquo

7

Pharmacokinetics

Distribution

How is a drug distributed to the various fluid

compartments or tissues of the body

Plasma vs interstitial vs intracellular fluid

Adipose tissue vs lean tissues

Distribution is unequal amp distinct for each drug

ldquoVolume of distributionrdquo ndash theoretical volume a drug

would occupy if it were present in all compartments at

the same concentration as in the plasma

8

ldquoVolume of distributionrdquo is based MOSTLY onhellip

A) how much blood a patient has

B) how much drug was in a particular pill

C) how soluble a particular drug is in lipids

vs water

9

True or false The volume of distribution of a

drug may be larger than the total volume of the

patient

A) True

B) False

10

Pharmacokinetics

Metabolism

Is a drug chemically modified by the body If sohellip

Are the metabolites more or less active

Are the metabolites safer or more toxic

Can the metabolites be more easily excreted

Which tissues metabolize the drug

What enzyme systems or other reactions are used to

metabolize the drug

11

Pharmacokinetics

Metabolism

Phase I reactions

Liver

Cytochrome P450 (cyp450) enzymes

Eg Oxidation reduction hydrolysis (de)cyclization

Often produce active metabolites

First-pass effect

Phase II reactions

Liver and kidney

Addition of polar functional groups

Usually produce non-functional metabolites for excretion

Eg Glucuronidation glutathione conjugation

12

Pharmacokinetics

Metabolic clearance of drugs hormones

Metabolized into non-active form

Bound to cells amp degraded recycled

Excreted

These processes are not mutually exclusive

Drugs can be metabolized to inactive metabolites then

excreted

13

Pharmacokinetics

Metabolic clearance of drugs hormones

Rate of removal depends on solubility

Peptide hormones

Degraded in blood or tissues

Cleared rapidly (seconds to minutes)

Steroid hormones

Bounds to plasma proteins

Cleared slowly (minutes to days)

14

Drug Hormone Clearance

Metabolic clearance rate (MCR)

Rate of disappearance from plasma

Infuse at increasing rate until plasma concentration

reaches steady-state

Infusion rate = rate of disappearance

Concentration of hormone in plasma

15

MCR (mLmin) = rate of hormone disappearance (ngmin)

Concentration of hormone (ngmL)

Pharmacokinetics

Excretion Elimination

Urine

Feces

Breath

Skin

16

Metabolic clearance rate refers tohellip

A) How fast kidneys produce urine

B) How much drug is filtered by the kidneys

C) A theoretical volume of plasma that can be

100 cleared of drug in a particular amount of

time

17

Pharmacodynamics

ldquoWhat a drug does to the bodyrdquo

Drug (ligand)-receptor interactions

Receptor types ndash membrane vs intracellular

Enzymes amp structural proteins can act as drug ldquoreceptorsrdquo

Signal transduction pathways

Enzyme-based biochemical pathways

18

L + R L R

Pharmacodynamics

Potential actionsroles of drugs

Agonist

Stimulating action vs depressing action

Full or partial agonist

Antagonist

Direct beneficial chemical reaction

Direct harmful chemical reaction

19

Pharmacodynamics

Dose-response curves

Increased [drug] increased effect

EC50 (effective dose yielding 50 max effect)

Curve shifts

Upward ndash higher efficacy

Leftward ndash higher potency (sensitivity)

20

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

For an orally-administered medication

absorption can only occur once which of these

other processes has occurred

A) Distribution

B) Metabolism

C) Liberation

D) Excretion

6

Why does the previous question include the

qualifer ldquoFor an orally-administered

medicationrdquo

7

Pharmacokinetics

Distribution

How is a drug distributed to the various fluid

compartments or tissues of the body

Plasma vs interstitial vs intracellular fluid

Adipose tissue vs lean tissues

Distribution is unequal amp distinct for each drug

ldquoVolume of distributionrdquo ndash theoretical volume a drug

would occupy if it were present in all compartments at

the same concentration as in the plasma

8

ldquoVolume of distributionrdquo is based MOSTLY onhellip

A) how much blood a patient has

B) how much drug was in a particular pill

C) how soluble a particular drug is in lipids

vs water

9

True or false The volume of distribution of a

drug may be larger than the total volume of the

patient

A) True

B) False

10

Pharmacokinetics

Metabolism

Is a drug chemically modified by the body If sohellip

Are the metabolites more or less active

Are the metabolites safer or more toxic

Can the metabolites be more easily excreted

Which tissues metabolize the drug

What enzyme systems or other reactions are used to

metabolize the drug

11

Pharmacokinetics

Metabolism

Phase I reactions

Liver

Cytochrome P450 (cyp450) enzymes

Eg Oxidation reduction hydrolysis (de)cyclization

Often produce active metabolites

First-pass effect

Phase II reactions

Liver and kidney

Addition of polar functional groups

Usually produce non-functional metabolites for excretion

Eg Glucuronidation glutathione conjugation

12

Pharmacokinetics

Metabolic clearance of drugs hormones

Metabolized into non-active form

Bound to cells amp degraded recycled

Excreted

These processes are not mutually exclusive

Drugs can be metabolized to inactive metabolites then

excreted

13

Pharmacokinetics

Metabolic clearance of drugs hormones

Rate of removal depends on solubility

Peptide hormones

Degraded in blood or tissues

Cleared rapidly (seconds to minutes)

Steroid hormones

Bounds to plasma proteins

Cleared slowly (minutes to days)

14

Drug Hormone Clearance

Metabolic clearance rate (MCR)

Rate of disappearance from plasma

Infuse at increasing rate until plasma concentration

reaches steady-state

Infusion rate = rate of disappearance

Concentration of hormone in plasma

15

MCR (mLmin) = rate of hormone disappearance (ngmin)

Concentration of hormone (ngmL)

Pharmacokinetics

Excretion Elimination

Urine

Feces

Breath

Skin

16

Metabolic clearance rate refers tohellip

A) How fast kidneys produce urine

B) How much drug is filtered by the kidneys

C) A theoretical volume of plasma that can be

100 cleared of drug in a particular amount of

time

17

Pharmacodynamics

ldquoWhat a drug does to the bodyrdquo

Drug (ligand)-receptor interactions

Receptor types ndash membrane vs intracellular

Enzymes amp structural proteins can act as drug ldquoreceptorsrdquo

Signal transduction pathways

Enzyme-based biochemical pathways

18

L + R L R

Pharmacodynamics

Potential actionsroles of drugs

Agonist

Stimulating action vs depressing action

Full or partial agonist

Antagonist

Direct beneficial chemical reaction

Direct harmful chemical reaction

19

Pharmacodynamics

Dose-response curves

Increased [drug] increased effect

EC50 (effective dose yielding 50 max effect)

Curve shifts

Upward ndash higher efficacy

Leftward ndash higher potency (sensitivity)

20

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Why does the previous question include the

qualifer ldquoFor an orally-administered

medicationrdquo

7

Pharmacokinetics

Distribution

How is a drug distributed to the various fluid

compartments or tissues of the body

Plasma vs interstitial vs intracellular fluid

Adipose tissue vs lean tissues

Distribution is unequal amp distinct for each drug

ldquoVolume of distributionrdquo ndash theoretical volume a drug

would occupy if it were present in all compartments at

the same concentration as in the plasma

8

ldquoVolume of distributionrdquo is based MOSTLY onhellip

A) how much blood a patient has

B) how much drug was in a particular pill

C) how soluble a particular drug is in lipids

vs water

9

True or false The volume of distribution of a

drug may be larger than the total volume of the

patient

A) True

B) False

10

Pharmacokinetics

Metabolism

Is a drug chemically modified by the body If sohellip

Are the metabolites more or less active

Are the metabolites safer or more toxic

Can the metabolites be more easily excreted

Which tissues metabolize the drug

What enzyme systems or other reactions are used to

metabolize the drug

11

Pharmacokinetics

Metabolism

Phase I reactions

Liver

Cytochrome P450 (cyp450) enzymes

Eg Oxidation reduction hydrolysis (de)cyclization

Often produce active metabolites

First-pass effect

Phase II reactions

Liver and kidney

Addition of polar functional groups

Usually produce non-functional metabolites for excretion

Eg Glucuronidation glutathione conjugation

12

Pharmacokinetics

Metabolic clearance of drugs hormones

Metabolized into non-active form

Bound to cells amp degraded recycled

Excreted

These processes are not mutually exclusive

Drugs can be metabolized to inactive metabolites then

excreted

13

Pharmacokinetics

Metabolic clearance of drugs hormones

Rate of removal depends on solubility

Peptide hormones

Degraded in blood or tissues

Cleared rapidly (seconds to minutes)

Steroid hormones

Bounds to plasma proteins

Cleared slowly (minutes to days)

14

Drug Hormone Clearance

Metabolic clearance rate (MCR)

Rate of disappearance from plasma

Infuse at increasing rate until plasma concentration

reaches steady-state

Infusion rate = rate of disappearance

Concentration of hormone in plasma

15

MCR (mLmin) = rate of hormone disappearance (ngmin)

Concentration of hormone (ngmL)

Pharmacokinetics

Excretion Elimination

Urine

Feces

Breath

Skin

16

Metabolic clearance rate refers tohellip

A) How fast kidneys produce urine

B) How much drug is filtered by the kidneys

C) A theoretical volume of plasma that can be

100 cleared of drug in a particular amount of

time

17

Pharmacodynamics

ldquoWhat a drug does to the bodyrdquo

Drug (ligand)-receptor interactions

Receptor types ndash membrane vs intracellular

Enzymes amp structural proteins can act as drug ldquoreceptorsrdquo

Signal transduction pathways

Enzyme-based biochemical pathways

18

L + R L R

Pharmacodynamics

Potential actionsroles of drugs

Agonist

Stimulating action vs depressing action

Full or partial agonist

Antagonist

Direct beneficial chemical reaction

Direct harmful chemical reaction

19

Pharmacodynamics

Dose-response curves

Increased [drug] increased effect

EC50 (effective dose yielding 50 max effect)

Curve shifts

Upward ndash higher efficacy

Leftward ndash higher potency (sensitivity)

20

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Pharmacokinetics

Distribution

How is a drug distributed to the various fluid

compartments or tissues of the body

Plasma vs interstitial vs intracellular fluid

Adipose tissue vs lean tissues

Distribution is unequal amp distinct for each drug

ldquoVolume of distributionrdquo ndash theoretical volume a drug

would occupy if it were present in all compartments at

the same concentration as in the plasma

8

ldquoVolume of distributionrdquo is based MOSTLY onhellip

A) how much blood a patient has

B) how much drug was in a particular pill

C) how soluble a particular drug is in lipids

vs water

9

True or false The volume of distribution of a

drug may be larger than the total volume of the

patient

A) True

B) False

10

Pharmacokinetics

Metabolism

Is a drug chemically modified by the body If sohellip

Are the metabolites more or less active

Are the metabolites safer or more toxic

Can the metabolites be more easily excreted

Which tissues metabolize the drug

What enzyme systems or other reactions are used to

metabolize the drug

11

Pharmacokinetics

Metabolism

Phase I reactions

Liver

Cytochrome P450 (cyp450) enzymes

Eg Oxidation reduction hydrolysis (de)cyclization

Often produce active metabolites

First-pass effect

Phase II reactions

Liver and kidney

Addition of polar functional groups

Usually produce non-functional metabolites for excretion

Eg Glucuronidation glutathione conjugation

12

Pharmacokinetics

Metabolic clearance of drugs hormones

Metabolized into non-active form

Bound to cells amp degraded recycled

Excreted

These processes are not mutually exclusive

Drugs can be metabolized to inactive metabolites then

excreted

13

Pharmacokinetics

Metabolic clearance of drugs hormones

Rate of removal depends on solubility

Peptide hormones

Degraded in blood or tissues

Cleared rapidly (seconds to minutes)

Steroid hormones

Bounds to plasma proteins

Cleared slowly (minutes to days)

14

Drug Hormone Clearance

Metabolic clearance rate (MCR)

Rate of disappearance from plasma

Infuse at increasing rate until plasma concentration

reaches steady-state

Infusion rate = rate of disappearance

Concentration of hormone in plasma

15

MCR (mLmin) = rate of hormone disappearance (ngmin)

Concentration of hormone (ngmL)

Pharmacokinetics

Excretion Elimination

Urine

Feces

Breath

Skin

16

Metabolic clearance rate refers tohellip

A) How fast kidneys produce urine

B) How much drug is filtered by the kidneys

C) A theoretical volume of plasma that can be

100 cleared of drug in a particular amount of

time

17

Pharmacodynamics

ldquoWhat a drug does to the bodyrdquo

Drug (ligand)-receptor interactions

Receptor types ndash membrane vs intracellular

Enzymes amp structural proteins can act as drug ldquoreceptorsrdquo

Signal transduction pathways

Enzyme-based biochemical pathways

18

L + R L R

Pharmacodynamics

Potential actionsroles of drugs

Agonist

Stimulating action vs depressing action

Full or partial agonist

Antagonist

Direct beneficial chemical reaction

Direct harmful chemical reaction

19

Pharmacodynamics

Dose-response curves

Increased [drug] increased effect

EC50 (effective dose yielding 50 max effect)

Curve shifts

Upward ndash higher efficacy

Leftward ndash higher potency (sensitivity)

20

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

ldquoVolume of distributionrdquo is based MOSTLY onhellip

A) how much blood a patient has

B) how much drug was in a particular pill

C) how soluble a particular drug is in lipids

vs water

9

True or false The volume of distribution of a

drug may be larger than the total volume of the

patient

A) True

B) False

10

Pharmacokinetics

Metabolism

Is a drug chemically modified by the body If sohellip

Are the metabolites more or less active

Are the metabolites safer or more toxic

Can the metabolites be more easily excreted

Which tissues metabolize the drug

What enzyme systems or other reactions are used to

metabolize the drug

11

Pharmacokinetics

Metabolism

Phase I reactions

Liver

Cytochrome P450 (cyp450) enzymes

Eg Oxidation reduction hydrolysis (de)cyclization

Often produce active metabolites

First-pass effect

Phase II reactions

Liver and kidney

Addition of polar functional groups

Usually produce non-functional metabolites for excretion

Eg Glucuronidation glutathione conjugation

12

Pharmacokinetics

Metabolic clearance of drugs hormones

Metabolized into non-active form

Bound to cells amp degraded recycled

Excreted

These processes are not mutually exclusive

Drugs can be metabolized to inactive metabolites then

excreted

13

Pharmacokinetics

Metabolic clearance of drugs hormones

Rate of removal depends on solubility

Peptide hormones

Degraded in blood or tissues

Cleared rapidly (seconds to minutes)

Steroid hormones

Bounds to plasma proteins

Cleared slowly (minutes to days)

14

Drug Hormone Clearance

Metabolic clearance rate (MCR)

Rate of disappearance from plasma

Infuse at increasing rate until plasma concentration

reaches steady-state

Infusion rate = rate of disappearance

Concentration of hormone in plasma

15

MCR (mLmin) = rate of hormone disappearance (ngmin)

Concentration of hormone (ngmL)

Pharmacokinetics

Excretion Elimination

Urine

Feces

Breath

Skin

16

Metabolic clearance rate refers tohellip

A) How fast kidneys produce urine

B) How much drug is filtered by the kidneys

C) A theoretical volume of plasma that can be

100 cleared of drug in a particular amount of

time

17

Pharmacodynamics

ldquoWhat a drug does to the bodyrdquo

Drug (ligand)-receptor interactions

Receptor types ndash membrane vs intracellular

Enzymes amp structural proteins can act as drug ldquoreceptorsrdquo

Signal transduction pathways

Enzyme-based biochemical pathways

18

L + R L R

Pharmacodynamics

Potential actionsroles of drugs

Agonist

Stimulating action vs depressing action

Full or partial agonist

Antagonist

Direct beneficial chemical reaction

Direct harmful chemical reaction

19

Pharmacodynamics

Dose-response curves

Increased [drug] increased effect

EC50 (effective dose yielding 50 max effect)

Curve shifts

Upward ndash higher efficacy

Leftward ndash higher potency (sensitivity)

20

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

True or false The volume of distribution of a

drug may be larger than the total volume of the

patient

A) True

B) False

10

Pharmacokinetics

Metabolism

Is a drug chemically modified by the body If sohellip

Are the metabolites more or less active

Are the metabolites safer or more toxic

Can the metabolites be more easily excreted

Which tissues metabolize the drug

What enzyme systems or other reactions are used to

metabolize the drug

11

Pharmacokinetics

Metabolism

Phase I reactions

Liver

Cytochrome P450 (cyp450) enzymes

Eg Oxidation reduction hydrolysis (de)cyclization

Often produce active metabolites

First-pass effect

Phase II reactions

Liver and kidney

Addition of polar functional groups

Usually produce non-functional metabolites for excretion

Eg Glucuronidation glutathione conjugation

12

Pharmacokinetics

Metabolic clearance of drugs hormones

Metabolized into non-active form

Bound to cells amp degraded recycled

Excreted

These processes are not mutually exclusive

Drugs can be metabolized to inactive metabolites then

excreted

13

Pharmacokinetics

Metabolic clearance of drugs hormones

Rate of removal depends on solubility

Peptide hormones

Degraded in blood or tissues

Cleared rapidly (seconds to minutes)

Steroid hormones

Bounds to plasma proteins

Cleared slowly (minutes to days)

14

Drug Hormone Clearance

Metabolic clearance rate (MCR)

Rate of disappearance from plasma

Infuse at increasing rate until plasma concentration

reaches steady-state

Infusion rate = rate of disappearance

Concentration of hormone in plasma

15

MCR (mLmin) = rate of hormone disappearance (ngmin)

Concentration of hormone (ngmL)

Pharmacokinetics

Excretion Elimination

Urine

Feces

Breath

Skin

16

Metabolic clearance rate refers tohellip

A) How fast kidneys produce urine

B) How much drug is filtered by the kidneys

C) A theoretical volume of plasma that can be

100 cleared of drug in a particular amount of

time

17

Pharmacodynamics

ldquoWhat a drug does to the bodyrdquo

Drug (ligand)-receptor interactions

Receptor types ndash membrane vs intracellular

Enzymes amp structural proteins can act as drug ldquoreceptorsrdquo

Signal transduction pathways

Enzyme-based biochemical pathways

18

L + R L R

Pharmacodynamics

Potential actionsroles of drugs

Agonist

Stimulating action vs depressing action

Full or partial agonist

Antagonist

Direct beneficial chemical reaction

Direct harmful chemical reaction

19

Pharmacodynamics

Dose-response curves

Increased [drug] increased effect

EC50 (effective dose yielding 50 max effect)

Curve shifts

Upward ndash higher efficacy

Leftward ndash higher potency (sensitivity)

20

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Pharmacokinetics

Metabolism

Is a drug chemically modified by the body If sohellip

Are the metabolites more or less active

Are the metabolites safer or more toxic

Can the metabolites be more easily excreted

Which tissues metabolize the drug

What enzyme systems or other reactions are used to

metabolize the drug

11

Pharmacokinetics

Metabolism

Phase I reactions

Liver

Cytochrome P450 (cyp450) enzymes

Eg Oxidation reduction hydrolysis (de)cyclization

Often produce active metabolites

First-pass effect

Phase II reactions

Liver and kidney

Addition of polar functional groups

Usually produce non-functional metabolites for excretion

Eg Glucuronidation glutathione conjugation

12

Pharmacokinetics

Metabolic clearance of drugs hormones

Metabolized into non-active form

Bound to cells amp degraded recycled

Excreted

These processes are not mutually exclusive

Drugs can be metabolized to inactive metabolites then

excreted

13

Pharmacokinetics

Metabolic clearance of drugs hormones

Rate of removal depends on solubility

Peptide hormones

Degraded in blood or tissues

Cleared rapidly (seconds to minutes)

Steroid hormones

Bounds to plasma proteins

Cleared slowly (minutes to days)

14

Drug Hormone Clearance

Metabolic clearance rate (MCR)

Rate of disappearance from plasma

Infuse at increasing rate until plasma concentration

reaches steady-state

Infusion rate = rate of disappearance

Concentration of hormone in plasma

15

MCR (mLmin) = rate of hormone disappearance (ngmin)

Concentration of hormone (ngmL)

Pharmacokinetics

Excretion Elimination

Urine

Feces

Breath

Skin

16

Metabolic clearance rate refers tohellip

A) How fast kidneys produce urine

B) How much drug is filtered by the kidneys

C) A theoretical volume of plasma that can be

100 cleared of drug in a particular amount of

time

17

Pharmacodynamics

ldquoWhat a drug does to the bodyrdquo

Drug (ligand)-receptor interactions

Receptor types ndash membrane vs intracellular

Enzymes amp structural proteins can act as drug ldquoreceptorsrdquo

Signal transduction pathways

Enzyme-based biochemical pathways

18

L + R L R

Pharmacodynamics

Potential actionsroles of drugs

Agonist

Stimulating action vs depressing action

Full or partial agonist

Antagonist

Direct beneficial chemical reaction

Direct harmful chemical reaction

19

Pharmacodynamics

Dose-response curves

Increased [drug] increased effect

EC50 (effective dose yielding 50 max effect)

Curve shifts

Upward ndash higher efficacy

Leftward ndash higher potency (sensitivity)

20

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Pharmacokinetics

Metabolism

Phase I reactions

Liver

Cytochrome P450 (cyp450) enzymes

Eg Oxidation reduction hydrolysis (de)cyclization

Often produce active metabolites

First-pass effect

Phase II reactions

Liver and kidney

Addition of polar functional groups

Usually produce non-functional metabolites for excretion

Eg Glucuronidation glutathione conjugation

12

Pharmacokinetics

Metabolic clearance of drugs hormones

Metabolized into non-active form

Bound to cells amp degraded recycled

Excreted

These processes are not mutually exclusive

Drugs can be metabolized to inactive metabolites then

excreted

13

Pharmacokinetics

Metabolic clearance of drugs hormones

Rate of removal depends on solubility

Peptide hormones

Degraded in blood or tissues

Cleared rapidly (seconds to minutes)

Steroid hormones

Bounds to plasma proteins

Cleared slowly (minutes to days)

14

Drug Hormone Clearance

Metabolic clearance rate (MCR)

Rate of disappearance from plasma

Infuse at increasing rate until plasma concentration

reaches steady-state

Infusion rate = rate of disappearance

Concentration of hormone in plasma

15

MCR (mLmin) = rate of hormone disappearance (ngmin)

Concentration of hormone (ngmL)

Pharmacokinetics

Excretion Elimination

Urine

Feces

Breath

Skin

16

Metabolic clearance rate refers tohellip

A) How fast kidneys produce urine

B) How much drug is filtered by the kidneys

C) A theoretical volume of plasma that can be

100 cleared of drug in a particular amount of

time

17

Pharmacodynamics

ldquoWhat a drug does to the bodyrdquo

Drug (ligand)-receptor interactions

Receptor types ndash membrane vs intracellular

Enzymes amp structural proteins can act as drug ldquoreceptorsrdquo

Signal transduction pathways

Enzyme-based biochemical pathways

18

L + R L R

Pharmacodynamics

Potential actionsroles of drugs

Agonist

Stimulating action vs depressing action

Full or partial agonist

Antagonist

Direct beneficial chemical reaction

Direct harmful chemical reaction

19

Pharmacodynamics

Dose-response curves

Increased [drug] increased effect

EC50 (effective dose yielding 50 max effect)

Curve shifts

Upward ndash higher efficacy

Leftward ndash higher potency (sensitivity)

20

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Pharmacokinetics

Metabolic clearance of drugs hormones

Metabolized into non-active form

Bound to cells amp degraded recycled

Excreted

These processes are not mutually exclusive

Drugs can be metabolized to inactive metabolites then

excreted

13

Pharmacokinetics

Metabolic clearance of drugs hormones

Rate of removal depends on solubility

Peptide hormones

Degraded in blood or tissues

Cleared rapidly (seconds to minutes)

Steroid hormones

Bounds to plasma proteins

Cleared slowly (minutes to days)

14

Drug Hormone Clearance

Metabolic clearance rate (MCR)

Rate of disappearance from plasma

Infuse at increasing rate until plasma concentration

reaches steady-state

Infusion rate = rate of disappearance

Concentration of hormone in plasma

15

MCR (mLmin) = rate of hormone disappearance (ngmin)

Concentration of hormone (ngmL)

Pharmacokinetics

Excretion Elimination

Urine

Feces

Breath

Skin

16

Metabolic clearance rate refers tohellip

A) How fast kidneys produce urine

B) How much drug is filtered by the kidneys

C) A theoretical volume of plasma that can be

100 cleared of drug in a particular amount of

time

17

Pharmacodynamics

ldquoWhat a drug does to the bodyrdquo

Drug (ligand)-receptor interactions

Receptor types ndash membrane vs intracellular

Enzymes amp structural proteins can act as drug ldquoreceptorsrdquo

Signal transduction pathways

Enzyme-based biochemical pathways

18

L + R L R

Pharmacodynamics

Potential actionsroles of drugs

Agonist

Stimulating action vs depressing action

Full or partial agonist

Antagonist

Direct beneficial chemical reaction

Direct harmful chemical reaction

19

Pharmacodynamics

Dose-response curves

Increased [drug] increased effect

EC50 (effective dose yielding 50 max effect)

Curve shifts

Upward ndash higher efficacy

Leftward ndash higher potency (sensitivity)

20

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Pharmacokinetics

Metabolic clearance of drugs hormones

Rate of removal depends on solubility

Peptide hormones

Degraded in blood or tissues

Cleared rapidly (seconds to minutes)

Steroid hormones

Bounds to plasma proteins

Cleared slowly (minutes to days)

14

Drug Hormone Clearance

Metabolic clearance rate (MCR)

Rate of disappearance from plasma

Infuse at increasing rate until plasma concentration

reaches steady-state

Infusion rate = rate of disappearance

Concentration of hormone in plasma

15

MCR (mLmin) = rate of hormone disappearance (ngmin)

Concentration of hormone (ngmL)

Pharmacokinetics

Excretion Elimination

Urine

Feces

Breath

Skin

16

Metabolic clearance rate refers tohellip

A) How fast kidneys produce urine

B) How much drug is filtered by the kidneys

C) A theoretical volume of plasma that can be

100 cleared of drug in a particular amount of

time

17

Pharmacodynamics

ldquoWhat a drug does to the bodyrdquo

Drug (ligand)-receptor interactions

Receptor types ndash membrane vs intracellular

Enzymes amp structural proteins can act as drug ldquoreceptorsrdquo

Signal transduction pathways

Enzyme-based biochemical pathways

18

L + R L R

Pharmacodynamics

Potential actionsroles of drugs

Agonist

Stimulating action vs depressing action

Full or partial agonist

Antagonist

Direct beneficial chemical reaction

Direct harmful chemical reaction

19

Pharmacodynamics

Dose-response curves

Increased [drug] increased effect

EC50 (effective dose yielding 50 max effect)

Curve shifts

Upward ndash higher efficacy

Leftward ndash higher potency (sensitivity)

20

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Drug Hormone Clearance

Metabolic clearance rate (MCR)

Rate of disappearance from plasma

Infuse at increasing rate until plasma concentration

reaches steady-state

Infusion rate = rate of disappearance

Concentration of hormone in plasma

15

MCR (mLmin) = rate of hormone disappearance (ngmin)

Concentration of hormone (ngmL)

Pharmacokinetics

Excretion Elimination

Urine

Feces

Breath

Skin

16

Metabolic clearance rate refers tohellip

A) How fast kidneys produce urine

B) How much drug is filtered by the kidneys

C) A theoretical volume of plasma that can be

100 cleared of drug in a particular amount of

time

17

Pharmacodynamics

ldquoWhat a drug does to the bodyrdquo

Drug (ligand)-receptor interactions

Receptor types ndash membrane vs intracellular

Enzymes amp structural proteins can act as drug ldquoreceptorsrdquo

Signal transduction pathways

Enzyme-based biochemical pathways

18

L + R L R

Pharmacodynamics

Potential actionsroles of drugs

Agonist

Stimulating action vs depressing action

Full or partial agonist

Antagonist

Direct beneficial chemical reaction

Direct harmful chemical reaction

19

Pharmacodynamics

Dose-response curves

Increased [drug] increased effect

EC50 (effective dose yielding 50 max effect)

Curve shifts

Upward ndash higher efficacy

Leftward ndash higher potency (sensitivity)

20

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Pharmacokinetics

Excretion Elimination

Urine

Feces

Breath

Skin

16

Metabolic clearance rate refers tohellip

A) How fast kidneys produce urine

B) How much drug is filtered by the kidneys

C) A theoretical volume of plasma that can be

100 cleared of drug in a particular amount of

time

17

Pharmacodynamics

ldquoWhat a drug does to the bodyrdquo

Drug (ligand)-receptor interactions

Receptor types ndash membrane vs intracellular

Enzymes amp structural proteins can act as drug ldquoreceptorsrdquo

Signal transduction pathways

Enzyme-based biochemical pathways

18

L + R L R

Pharmacodynamics

Potential actionsroles of drugs

Agonist

Stimulating action vs depressing action

Full or partial agonist

Antagonist

Direct beneficial chemical reaction

Direct harmful chemical reaction

19

Pharmacodynamics

Dose-response curves

Increased [drug] increased effect

EC50 (effective dose yielding 50 max effect)

Curve shifts

Upward ndash higher efficacy

Leftward ndash higher potency (sensitivity)

20

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Metabolic clearance rate refers tohellip

A) How fast kidneys produce urine

B) How much drug is filtered by the kidneys

C) A theoretical volume of plasma that can be

100 cleared of drug in a particular amount of

time

17

Pharmacodynamics

ldquoWhat a drug does to the bodyrdquo

Drug (ligand)-receptor interactions

Receptor types ndash membrane vs intracellular

Enzymes amp structural proteins can act as drug ldquoreceptorsrdquo

Signal transduction pathways

Enzyme-based biochemical pathways

18

L + R L R

Pharmacodynamics

Potential actionsroles of drugs

Agonist

Stimulating action vs depressing action

Full or partial agonist

Antagonist

Direct beneficial chemical reaction

Direct harmful chemical reaction

19

Pharmacodynamics

Dose-response curves

Increased [drug] increased effect

EC50 (effective dose yielding 50 max effect)

Curve shifts

Upward ndash higher efficacy

Leftward ndash higher potency (sensitivity)

20

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Pharmacodynamics

ldquoWhat a drug does to the bodyrdquo

Drug (ligand)-receptor interactions

Receptor types ndash membrane vs intracellular

Enzymes amp structural proteins can act as drug ldquoreceptorsrdquo

Signal transduction pathways

Enzyme-based biochemical pathways

18

L + R L R

Pharmacodynamics

Potential actionsroles of drugs

Agonist

Stimulating action vs depressing action

Full or partial agonist

Antagonist

Direct beneficial chemical reaction

Direct harmful chemical reaction

19

Pharmacodynamics

Dose-response curves

Increased [drug] increased effect

EC50 (effective dose yielding 50 max effect)

Curve shifts

Upward ndash higher efficacy

Leftward ndash higher potency (sensitivity)

20

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Pharmacodynamics

Potential actionsroles of drugs

Agonist

Stimulating action vs depressing action

Full or partial agonist

Antagonist

Direct beneficial chemical reaction

Direct harmful chemical reaction

19

Pharmacodynamics

Dose-response curves

Increased [drug] increased effect

EC50 (effective dose yielding 50 max effect)

Curve shifts

Upward ndash higher efficacy

Leftward ndash higher potency (sensitivity)

20

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Pharmacodynamics

Dose-response curves

Increased [drug] increased effect

EC50 (effective dose yielding 50 max effect)

Curve shifts

Upward ndash higher efficacy

Leftward ndash higher potency (sensitivity)

20

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Pharmacodynamics

21

0 50 100 1500

25

50

75

100

125

LPO AdLib (n=15)

NPO AdLib (n=11)

NPO FR (n=5)

LPO FR (n=8)

KCl (mM)

Ten

sio

n (

mN

mm

)

01 1 10 10

010

00

1000

0

1000

00

0

20

40

60

80

100

120

140

160

NorEpi (nM)

Ten

sio

n (

mN

mm

)

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

A leftward shift of a dose-response curve

indicates that a systemhellip

A) is more senstive to a particular ligand

B) is less sensitive to a particular ligand

C) has a higher concentration of ligand

C) has a lower concentration of ligand

22

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Pharmacodynamics

Beneficial actions of drugs must be

weighed against unintended harmful

effects

Side effects

Therapeutic window

The range of doses which provide benefits

without producing toxicity

Varies for each drug and may vary between

individuals

23

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

True or false Medications with side effects are

not approved by the FDA

A) True

B) False

24

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Measurement of Hormone Concentration

Radioimmunoassay

Use specific antibody to detect hormone

Mix radioactive ldquotracerrdquo hormone antibody and

sample together

Competitive assay [Ab] is lower than combined

tracer and endogenous [hormone]

Remove all unbound mixture leaving Ab bound

to either tracer or endogenous hormone

High [endogenous hormone] low tracer

Low [endogenous hormone] high tracer

25

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Measurement of Hormone Concentration

Radioimmunoassay

Measure radioactivity of remaining sample

compare to standard curve

Fig 74-9 26

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Measurement of Hormone Concentration

ELISA (enzyme-linked immunosorbent

assay)

Coat sample wells with antibody

Add test sample

Add antibody conjugated to enzyme

Add enzyme substrate

Color development

indicates presence of

test substance

concentration

determined by dilution

factor amp comparison to

standard curve 27

Fig 74-10

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

Mouse ICAM-1 ELISA Standard Curve

httpswwwthermofishercomordercatalogproductEMICAM1

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

What kind of molecule is used as a ldquomolecular

toolrdquo to measure how much of a particular

substance a sample has

A) Protein

B) Receptor

C) DNA

D) Antibody

29

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30

RIA standard curves have a negative slope

ELISA standard curves are positive Why are

they different

30