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Validation & Biomanufacturing

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What is Validation

Validation An Essential Part of GMPs!

Validation is the scientific study of a system

To prove that the facility/system/equipment/method is consistently doing

what it is supposed to do (i.e., that the process is under control).

We want to make decisions based on good science and not hunches andassumptions!

To determine the process variables and acceptable limits for these variables,

and to set-up appropriate in-process controls.

Is it ok if the wash from a chromatography column is pH 6.8 vs. 7.0 ?

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FDA definition of validation

Validation is a process ofdemonstrating, through

documented evidence, that a process, procedure,method, piece of equipment, or facility will

consistentlyproduce a product or result that meets

predetermined specifications and quality

attributes.

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Quality Attributes Remember these?Identity

21 CFR211.84 (d) at least one test shall be conducted to verify the identity of each component of a drug product.

Chemical, biological, Immunological Raw materials, In-process intermediates, final products.

Safety

21 CFR 600.3 (p) safety as the relative freedom from harmful effect to persons affected, directly or indirectly, by aproduct when prudently administered, taking into consideration the character of the product in relationship to thecondition of the recipient at the time.

Activity of active ingredients

Activity of the excipients or additives

Activity of process related impurities

Efficacy

Effectiveness of the product in achieving its medicinal purpose (therapeutic, prophylactic, diagnostic). Gathered atphase II and Phase III trials.

Potency

21 CFR 600.3 (s) specific ability or capacity of the product, as indicated by its appropriate laboratory tests or byadequately controlled clinical data obtained through the administration of the product in the manner indicated toeffect the given result.

Purity

21 CFR 600.3 (r) relative freedom from extraneous matters in the finished product, whether or not harmful to therecipient or deleterious to the product.

Cleaning Procedures

Stability

21 CFR211.137 (a) to assure that a drug product meets applicable standards of identity, quality, and purity at thetime of use; it shall bear an expiration date determined by stability testing. Drugs may use accelerated time studies,biologics must use real time studies.

Consistency

The ability of the product and/or process to reliably possess specified quality attributes on an ongoing basis. 3consecutive batches of product meeting predetermined specifications is accepted as proof that a process isconsistent. However, in NDA data from up to twenty batches may be submitted.

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Biomanufacturing

Biomanufacturing is a complex process involving multiple unit

operations many of which are critical to insuring patient safety and

product efficacy

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InnoculumSeed

Fermentati

on

ProductionFermentati

on

Harvest

Ultrafiltrati

on

1

Chrom. 11

Ultrafiltrati

on

2

Chrom.

2

Viral

Filtration

Chrom.

3

Ultrafiltrati

on

3

Final

Formulation/

Sterile Filtration

Sterile Fill

UPSTREAM

DOWN-

STREAM

VIRAL

NON-VIRAL

Block Flow Diagram of a typical Production Process

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Historical Basis for Validation

Assumptions concerning virus inactivation resulted in tendeaths and 200 children becoming paralyzed, from asupposedly inactivated polio vaccine.

Assumptions about sterilization caused severe infectionsamong burn victims given supposedly sterile solutions.

Validation eliminates assumptions and relies on

experimental proof!

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Quality by Design

A central concept in quality is that quality can not be tested for.

Quality must be designed and built into the production process.

Requires careful attention to raw material specifications, in

process material specifications, and final product specifications.

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Validation and Quality

Validating the performance of unit operations, analytical

methods, and critical process points (sterilization, viral

inactivation, cleaning procedures) is essential in insuring that

the process generates a quality product.

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Validation in Biomanufacturing

Validation does not replace testing, but it does reduce thetesting burden for raw materials, in-process materials, and

final product

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Validation in Biomanufacturing

Validation itself is a process that evolves with the

product.

Validation requirements for production of pre-clinicalmaterial less stringent then for phase III clinical

material.

Critical operations must be validated: For example:

raw materials, analytical methods, viral clearance,sterilization, cleaning.

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Validation in Biomanufacturing

A fully validated process is locked in

Any change outside of the validated space invalidates process

Change must be evaluated for effect on patient safety andproduct efficacy-Change control !

Validated Production Process

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Less

More

Difficulty in Changing and Binding Authority

LAWS

Regulation

Guidance

Less

More

LAWS

Regulation

Guidance

Scientific Content & Detail about Implementation

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Regulatory requirement for validation

21 CFR211 Subpart H- Holding and Distribution

211.165 Testing and release for distribution

Requires that the accuracy, sensitivity, specificity, andreproducibility of test methods employed by the firmshall be established and documented. Such validationand documentation may be accomplished in accordance

with 21 CFR 211.194 (a)(2)

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Regulatory requirement for validation

Sec. 211.113 Control of microbiological contamination.

(a) Appropriate written procedures, designed to prevent

objectionable microorganisms in drug products not required to besterile, shall be established and followed.

(b) Appropriate written procedures, designed to preventmicrobiological contamination of drug products purporting to besterile, shall be established and followed. Such procedures shallinclude validation of any sterilization process.

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What does validation of any

sterilization process mean ? What parameters are critical to sterilization?

Temperatures, pressures, time, pore size (filtration), radiationdosage, chemical concentration.

Must demonstrate that your autoclave reaches the temperatures,pressures, and times necessary for sterilization.

Must demonstrate that items representing real world samples achievethose conditions ( 20 ft of1 hose; a 20 L carboy; a 500 ml bottle).

Must challenge with worse case scenario (may take place in pilotplant if scalability demonstrated).

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Regulatory guidance on validation

Guideline on General Principals of Process Validationhttp://www.fda.gov/cder/guidance/pv.htm

Guidance for Industry: For the Submission Documentation for SterilizationProcess Validation in Applications for Human and Veterinary DrugProducts. CDER CVM November1994.www.fda.gov/CDER/GUIDANCE/cmc2.pdf

Working Party on Control of Medicines and Inspections Final Version of Annex 15 to the EU Guide to Good Manufacturing Practice

Title: Qualification and validation

http://pharmacos.eudra.org/F2/eudralex/vol-4/pdfs-en/v4an15.pdf

ICH Q7a Section 12 on validation

http://www.fda.gov/cder/meeting/ICH_Q7A/index.htm

A WHO guide to good manufacturing practice (GMP) requirements. Part 2:Validation

Chaloner-Larsson, G., Anderson, R., and Egan, A. 1997. World HealthOrganization, Geneva.

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Critical Operations in

Biomanufacturing

Some operations are more critical than others.

Viral filtration, sterilization, cleaning, analytical methods.

These operations will require greater validation efforts then less

critical operations (media blending).

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Testing

Usually done by the Quality Control Laboratory

CFR requires that quality unit be under independent

supervision and report directly to senior management

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Quality Assurance

Reviews records from quality control and production departments

Verifies that all specifications and production operations met /performed

Investigations necessary for any deviations

Root cause

Affect on quality

Corrective action (CAPA)

Approves final release of product

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Designing Quality into the Product Design of production process and specifications all contribute

to a quality product:

Absence of contamination

Clean rooms, closed systems, use ofBSC for criticaloperations.

Purity

Separation process (chromatography) designed toremove potential contaminants

Viral purification / inactivation

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Validation Plan

Organizations must define an approach towards

validation

What is to be validated

How is it to be validated

Who is to validate it

Who is to approve the validation When it must be revalidated

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ValidationExamples of individual

systems subject to validation:

HVAC systems

Autoclaves

pH meters

Depyrogenation Ovens

Lyopholyzers

Centrifuges

Steam generatorsWater systems

Compressed air systems

Vacuum systems

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Validation Plan

Regulatory agencies (FDA, EMEA, WHO, etc) identify minimumcomponents of validation.

Industry standards (the c in cGMP) can increase validationrequirements.

New & Novel processes / equipment require greater scrutiny thenestablished processes / equipment.

Validation requirements increase as a product moves throughdevelopment (phase I, phase II, phase III).

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A prospective validation study

IQ

OQ

Calibration

PQ protocol

approval

PQ protocolexecution

Data

Analysis

ValidationReport

Approve

Conclusions

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Developmental studies

Experiments designed to explore and define the limits of the system to be

validated

Sterilization developmental studies may focus on

worst case orhard to sterilize items

Cleaning developmental studies may focus on worst case or hard to

clean items

Analytical methods may focus on defining the limits of the procedure

(range, recovery, etc)

Developmental studies then used to develop validation protocols and

refine SOPs

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Validation Protocol

Specific protocols (SOPs) that provide detailed information on what is tobe validated.

Validation Protocols consist of: A description of the process, equipment, or method to be validated.

A description of the validation method.

A description of the sampling procedure including the kind andnumber of samples.

Acceptance criteria for test results. Schedule or criteria for revalidation.

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Example of a protocol for the

IQ component of validating a

pH meter

As with all other SOPs this

document will contain an

Objective, scope, and responsibility

Section.

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Validation Protocol

Validation Protocols may consist of multiple SOPs each

describing specific steps in the validation process

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Critical Systems

How critical is the system being validated to final product quality?

Media blending systems for cell growth vs. final fill & finishoperations

Demonstrating that the device which fills, labels, and caps thefinal product will require more extensive validation then the

blenders used to prepare media for bioreactors.

Validation of complex devices may take years!

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Validation

Proceeds in stages with new facilities / equipment.

Planning for validation should start with the design

process.

Leaving validation to the last minute is asking for

trouble.

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Stages of Validation

Starts with Design & Receipt:

Does the equipment meet the needs (is the autoclave bigenough?)

Do you have the manuals, spare parts, can you plug it in?

Is it installed properly (drain lines, vents, etc) Does it work?

Does the autoclave reach the necessary temp. and pressure?

Can the autoclave sterilize your equipment (worse casesituation)?

How does it work in the manufacturing process? Can it handle production quantities?

Will failure compromise product quality?

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IQ, OQ, PQ ?

Installation Qualification (IQ)

A process used to document that the piece of equipment was supplied andinstalled properly and that appropriate utilities, i.e., electrical, steam, gas,etc. are available to operate the equipment according to the manufacturersspecifications.

Operational Qualification (OQ)

A process designed to supply the documented evidence that a piece ofequipment operates as it is intended through all anticipated operationalranges.

Performance (Process) Qualification (PQ)Verifies that a process / piece of equipment performs as it is intended to inthe manufacturing process and produces product (in process or final)meeting predetermined specifications.

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Typical information in an IQ protocol

Name and description of equipment, including model numbers

Identification, including model and serial numbers

Location of the equipment Any utility requirements, i.e. electrical voltage, steam or water

pressure, etc.

Any safety features of the equipment, including alarms, interlocks,or relief valves.

That all documentation, including manufacturers contact

information, spare parts inventory, operational manual, andinstallation drawings are available on site.

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Typical OQ Protocol Components

Objective

Responsibility Equipment required (Calibration verification & Traceability)

SOP(s) used

Equipment Identification

Parameters measured (Specifications)

Documentation

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Validation

Ideally validation takes place prior to actual production runs,however in some cases validation may take place as productis produced, or past production runs may be used to providevalidation data.

ProspectiveValidation

ConcurrentValidation

Retrospective Validation

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Revalidation

Is the initial validation of a piece of equipment the end?

No!

Periodic revalidation may be necessary depending on the criticalityof the equipment

Changes need to be evaluated for their impact on validation

Deviations from specifications may require revalidation

Revalidation spelled out in MasterValidation Plan

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Change Control

Must assess impact of changes on FDA compliance and validation

state.

Change control is a formal process defined in company SOP on howprocess/equipment changes are evaluated.

Any change that takes place outside the change control process canjeopardize product quality (patient safety).

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References Pharmaceutical Manufacturers Associations (Pharmaceutical Research and Manufacturers of America) Validation

Advisory Committee ProcessValidation Concepts for Drug Products Pharmaceutical Technology, September1985 p 82.

Bismuth, G. CleaningValidation: A Practical Approach. CRC Press, 2000. ISBN 1574911082. Pharmaceutical ProcessValidation, 3rd Ed. Edited by Robert Nash and Alfred Wachter, Marcel Decker, 2003. ISBN

082470838-5

Validation of Pharmaceutical Processes:Sterile Products. 1998. 2nd Edition.Edited by Frederick J. Carlton and JamesAgalloco. Marcel Decker, 1998. ISBN 0824793846.

Validation Standard Operating Procedures:A step by Step Guide for Achieving Compliance in the Pharmaceutical, MedicalDevice, and Biotech Industries, Syed Imtiaz Haider, St. Lucie Press, 2002. ISBN 1574443313.

Good Manufacturing Practices for Pharmaceuticals:A Plan for Total Quality Control From Manufacturer to Consumer,Sidney J. Willig. Marcel Decker, 2000. ISBN 0824704258.

Voss, J. Cleaning and Cleaning Validation: A Biotechnology Perspective. CRC Press, 1995. ISBN 0939459507.

LeBlanc, D.A. 2000.Validated Cleaning Technologies for Pharmaceutical Manufacturing. CRC Press. ISBN 1574911163. Cloud, P. 1998. PharmaceuticalEquipmentValidation: The Ultimate Qualification Guidebook. CRC Press. ISBN

1574910795.

Juran, Quality Control Handbook, 4th Edition., McGraw-Hill, 1988.

DeSain C, Sutton C. (1995). Process development that supports process validation. Pharmaceutical Technology19 (Oct.):130-136, 1995.

Garcia T, Wilkinson S, Scott J. The development of a blend-sampling technique to assess the uniformity of a powdermixture. DrugDevelopment and Industrial Pharmacy27(4):297-307,2001.

Chaloner-Larsson, G., Anderson, R.,Egan, A. 1997. A WHO guide to good manufacturing practice (GMP) requirementsPart 2:Validation . World Health Organization, Geneva. www.who.int/vaccines-documents/DocsPDF/www9666.pdf

Accessed on October2nd, 2006. Brown, F. 1993. Review of accidents caused by incomplete inactivation of viruses. Dev. Biol. Stand. 81: 103-7

Nathanson, N. and Langmuir, A.D. 1995. The Cutter incident. Poliomyelitis following formaldehyde-inactivated poliovirusvaccination in the United States during the Spring of1955. II. Relationship of poliomyelitis to Cutter vaccine. 1963.Am. J.Epidemiol.142:109-40.

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Autoclave Validation

IQ-

Design specifications meet users needs

Proper installation, utilities, manuals, spare parts

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The DQ, IQ, OQ process

insures that this autoclave

will meet the needs of the

manufacturing group.

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Sample Format forInstallationQualification of an autoclave.

Courtesy of WHO. Chaloner-Larsson, G.,Anderson, R.,Egan, A. 1997. A WHO guide to good

manufacturing practice (GMP) requirements Part 2:Validation .

World Health Organization, Geneva. www.who.int/vaccines-documents/DocsPDF/www9666.pdf

Accessed on October 2nd, 2006.

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Autoclave Validation

OQ

Does it operate properly Does it reach the specified temperature and

pressure

Do timers work

Does the operator interface panel work

Are safety interlocks functional

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Sample Format forOperational

Qualification of an autoclave.

Courtesy of WHO. Chaloner-Larsson, G.,Anderson, R.,Egan, A. 1997. A WHO guide to good

manufacturing practice (GMP) requirements Part 2: Validation .World Health Organization, Geneva

. www.who.int/vaccines-documents/DocsPDF/www9666.pdfAccessed on October 2nd, 2006.

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Monitoring Temperature

Figure 2: A validator, used in the

operational qualification of an

autoclave. The validator is

attached to individual

thermocouples by wires comingfrom the rear of the instrument

(arrow). Tha validator has been

previously calibrated and the data

gathered from the thermocouples

will be logged on the laptop

computer. The software on thecomputer is also subject to

validation requirements.

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Figure 4. The validator is

attached to individual

thermocouples (TC) by thin

wires that pass through the

wall of the autoclavethrough a specially

designed port (arrow). This

picture shows the back side

of the autoclave. The

validator is out of view at

the lower left.

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Figure 5. The inside of the

autoclave showing the maze

of wiring connecting the

individual TCs to the

validator. The port through

which the wires pass is

visible in the middle left of

the picture. The individual

TCs will be placed in various

areas of the autoclave or

equipment being autoclavedto generate a thermal map of

the interior of the autoclave.

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Figure 3. Output from the

validator. The temperature at

each connected thermocouple is

displayed. Accumulated lethality

(F0) may also be displayed.Notice how some TChave failed

and will not record a

temperature. Accounting for TC

failure is necessary to keep from

having to repeat a study.

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Developmental Studies

Developmental studies are

used to identify hard to

autoclave items and totest if item preparation has

an effect on ability to be

sterilized

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Developmental Studies

Every little nook and cranny

Needs to be assessed ! We

have to prove that the inside

of the pipe reaches sufficient

temperature, for a long

enough time to insure sterility!

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Developmental Studies

Does bagging make a difference?

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Cleaning Validation

Validating the cleaning cycle

on a loaded dishwasher.

Notice the various pipes andparts with narrow openings.

Identification of hard to clean

and easy to clean areas starts

with developmental studies

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How do we validate a dishwasher ?

How do we identify hard to clean & easy to clean items?

How do we test to see if they are cleaned?

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Going over the documentation

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Cleaning Validation

Sampling, documenting, and

verifying is a labor intensive

process

Why is cleaning considered a

critical process?

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Sampling

Figure 8. Swabbing is

commonly used to sample the

surface of cleaned materials.

The swab is then placed in a

sample vial and sent to theQuality Control lab for

analysis. Proper technique is

essential in order to evaluate

the effectiveness of cleaning

techniques. Even so,

swabbing results are typically

corrected for knowndeficiencies in recovery.

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Cleaning Validation

Figure 9. Analysis of rinse water

for residual cleaning agents orprocess materials is an essential

component of cleaning validation.

Insuring that the sample is not

contaminated requires vigilance

and properly following the

relevantSOPs.

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Analytical methods validation

Considered a critical step in the manufacturing process

Requirements for validated analytical methods explicitly written intothe CFRs

211.165 Testing and release for distribution

Requires that the accuracy, sensitivity, specificity, and reproducibility oftest methods employed by the firm shall be established and documented.Such validation and documentation may be accomplished in accordancewith 21 CFR211.194 (a)(2)

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Testing For Identity

Requires the development of validated analytical methods that can

determine identity. Chemical Tests:

Is the molecule chemically what it is supposed to be?

Biological Activity Tests:

Does the molecules have the biologic activity that it is supposed tohave?

Immunogenic Tests:

Is the molecule immunogenic (allergic)?

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Identity

21 CFR requires testing of raw materials:

Raw materials quarantined until identity verified

Raw materials must meet predetermined specifications

Vendors (and alternates) specified in BLA (NDA)

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Identity

21 CFR requires testing of in-process materials:

Product from bioreactor / fermentor Product from purification steps

Waste products from above

Must meet specifications, if not - stop the

process to investigate take corrective action

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Regulatory guidance on validation

Guideline on General Principals of Process Validationhttp://www.fda.gov/cder/guidance/pv.htm

Guidance for Industry: For the Submission Documentation for SterilizationProcess Validation in Applications for Human and Veterinary DrugProducts. CDER CVM November1994.www.fda.gov/CDER/GUIDANCE/cmc2.pdf

Working Party on Control of Medicines and Inspections

Final Version of Annex 15 to the EU Guide to Good Manufacturing Practice

Title: Qualification and validation

http://pharmacos.eudra.org/F2/eudralex/vol-4/pdfs-en/v4an15.pdf

ICH Q7a Section 12 on validation

http://www.fda.gov/cder/meeting/ICH_Q7A/index.htm

A WHO guide to good manufacturing practice (GMP) requirements. Part 2:Validation

Chaloner-Larsson, G., Anderson, R., and Egan, A. 1997. World HealthOrganization, Geneva.

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Validation Protocol

Specific protocol based on developmental studies

Protocol is written, reviewed and approved Protocol is executed

Report written and approved

System is validated

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Analytical Methods Validation

B

eing a critical component of productionprocess analytical methods must be validated

Raw material testing

In process materials

Final product specifications

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What must be demonstrated

Selectivity (specificity)

Accuracy

Precision Linear Range

Limit of detection (LOD)

Limit of quantification (LOQ or LLOQ)

Robustness

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Validated methods

USP NF (United States Pharmacopeia National Formulary) contains

validated analytical methods

Use of a USP method does not eliminate the organizations

obligation to demonstrate that the method performs adequetly

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Selectivity (specificity)

Does the analytical method detect the component it is supposed

to detect? Cross reactivity in antibody based methods

Demonstrate specificity by conducting analytical method on

materials that may mimic analyte of interest

Looking forfalse positives

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Accuracy

Ability of analytical method to accurately determine the

presence and amount of the analyte of interest Typically done by analyzing a traceable standard

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Linear Range

Must define the linear range of a method

Assay may have multiple linear ranges

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Precision

How much variability does the assay exhibit when analyzingthe same sample

Typically demonstrated by analyzing multiple aliquots of a

homogenous sample

Acceptance criteria will depend on the assay and the

material being assayed (2-20% RSD)

Typically expressed as % RSD (relative standard deviation)

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Limit ofDetection (LOD)

Lowest level at which method can detect analyte

Results reported as less than LOD

Based on signal to noise specification (10:1, 20:1)

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Limit ofQuantification (LOQ)

Lowest level at which method can accurately quantify analyte

Based on signal-to-noise ratio specification (10:1, 20:1)

and precision specification

Precision and LOQ related

Lower LOQ will typically result in lower precision

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Bradford Assayfor total protein

Well known colorimetric assay that relies on the binding of

Commassie G-250 dye to the proteins in an acidic solution

Dye binding proportional to number of positive charges in

protein

Proteins >3000 dal not detected

Simple, quick, wide range, few interfering agents

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BradfordA

ssay

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Disadvantages

Incompatability with surfactants Staining of glass and quartz cuvettes

Use disposable polystyrene cuvettes

Or wash with strong detergents and methanol

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Validating the Bradford

Selectivity (specificity)

Accuracy

Recovery

Precision

Linear Range

Limit of detection (LOD)

Limit of quantification (LOQ or LLOQ) Robustness

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Some Questions

A valve used to transfer material from a holding tank to the purification suitejams closed. You have a spare valve that is an identical model. Can you changethis valve with the spare and continue operations? What if the valve is from adifferent manufacturer?

You notice that your autoclave loading plan leaves room for additional material.

Realizing that increasing that amount of material in the autoclave will shortenthe turn around time for the production line you contemplate increasing theamount of material loaded into the autoclave then specified by the loading plan.What should you do? What will be required to implement this change?

An SOP for calibration of a pH meter calls for a two point calibration at pH 4and pH 7. You notice that a single point calibration at pH 7 produces the same

result from pH measurements of your buffer solutions and allows you to take alonger break. Is it Ok to do the one point calibration when the SOP calls for atwo point calibration? How would you go about changing the SOP to allow for aone point calibration?

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What documents would provide information concerning the make and

model of a particular valve used to regulate the transfer of material from a

holding tank to the purification suite?

Your supervisor is concerned that the fermentation vessel is not providing

sufficient aeration of the culture to get optimal growth and suggests

installing a different kind of baffle in the vessel. How would you

demonstrate that this change has no effect on product quality?