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Meso Scale Discovery ®
Bioprocess Applications
Meso Scale Discovery Bioprocess Applications
MSD offers a diverse product line of assays and
kits for application in Bioprocess. Kits for
measuring host cell protein contamination and
contamination by other impurities can replace
more cumbersome ELISA, western blot or HPLC
methods. MSD assays can measure antibody
expression levels in production or anti-drug
antibodies in immunogenicity testing. Ultra-
sensitive cytokine and phosphoprotein kits can
enable functional cell based measurements and
general potency assays. MSD’s instruments are
well-suited for regulated environments. MSD
DISCOVERY WORKBENCH® 3.0 Software is
21 CFR Part II compliant and we offer
comprehensive packages for instrument
validation.
Kits & Assays for HCP Contamination
Kits & Assays for Impurity & Residual Contamination
Kits for Multiplexed Assays for Contamination
Assays for Measuring Antibody Expression
Functional Cell-Based Assays
Cytokine & Phosphoprotein Potency Assays
Cell-Based Assays for Antibody Screening
Immunogenicity Assays
Meso Scale DiscoveryA division of Meso Scale Diagnostics, LLC.
9238 Gaither Road Gaithersburg, MD USA 20877Phone: 240.631.2522 (x4685 for customer service)
www.mesoscale.com
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Electrochemiluminescence Features:Minimal background signals and high signal to back-
ground ratios - the stimulation mechanism (electricity) is decoupled from the signal (light)
Proximity - only labels bound near the electrode surface are detected, enabling non-washed assays
Flexibility - labels are stable, non-radioactive, and are conveniently conjugated to biological molecules
Emission at ~620 nm - eliminating problems with color quenching
Signal amplification - multiple excitation cycles of each label enhance light levels and improve sensitivity
MSD’s electrochemiluminescence detection technology uses SULFO-TAGTM labels that emit light upon electrochemical stimulation initiated at the electrode surfaces of MULTI-ARRAY® and MULTI-SPOT® microplates.
1 2
A
BMULTI-ARRAY Plate
Ru(bpy)33+Ru(bpy)3
2+
*Ru(bpy)32+
TPA TPA.+
TPA.-H+
e-e-
LIGHT
LuminescenceEmitting Light
Chemi-Chemical Energy
Electro-Electrochemically Initiated
Measured signal is light
Working Electrode DielectricCounter Electrode
Simultaneous measure-ment of analytes in one well. Capture antibodies are arrayed on the patterned working electrode. Analytes are detected with antibodies labeled with SULFO-TAG reagent.
Dielectric
Counter electrode
Analyte 1
Analyte 2
Analyte 3
Analyte 4
MULTI-ARRAY and MULTI-SPOT Features: Flexible surface coatings to suit most any biology
Carbon electrode plate surface has 10X greater binding capacity than polystyrene
High density arrays for high content screening and high throughput multiplexing of biomarkers
Custom surface coatings and patterns
MULTI-SPOT Assay
MSD Technology
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Kits and Assays for Host Cell Protein (HCP) Contamination
Contaminants from host cells are typically measured by ELISAs or western blots, which suffer long assay times, complex protocols, and small dilutional linear range. MSD offers assays with simple protocols that are usually completed in less than 4 hours. Most assays require few or no wash steps. MSD assays are easy to develop: HCP assays for CHO, E.coli, NS/0, and HEK293 cell lines have been developed with both MSD antibodies and customer antibodies.
Using the right antibodies is critical in contamination assays. MSD offers you the choice of ready-made kits with commercial antibodies or custom kits that incorporate your process-specific antibodies. Our assay development tools let you build assays without modifying your reagents. Whether you prefer direct immobilization or binding through an intermediate (strepavidin/biotin, anti-species antibodies, Protein A), MSD has the tools you need.
Sign
al
Concentration (pg/mL)**
10,000
10 10,000 100,000
1,000
100 1,000
MSD vs ELISA-CHO HCP Assay
Abs
orba
nce
(492
nm
-405
nm
)0.8
0.65
0.55
0.45
0.35
0.25
0.3
Electrode Surface
MSD Kits for General HCP Contamination Assays
MSD offers a preconfigured kit for CHO HCP contamination assays. This kit includes plates pre-coated with capture antibody, CHO standards, diluents, and pre-labeled detection antibodies. The MSD assay is 100 times more sensitive than the comparable ELISA (with the same antibodies). The wide dynamic range (3.5 logs) expands the working range and eliminates the need for large dilutions. The MSD protocol has a single wash step and a total assay time of 4 hours.
Host cellprotein
SULFO-TAG-labeled anti-CHOHCP antibody
Anti-CHO HCPcapture antibody
CHO Host Cell Protein Kit (96-well plates)
MSD (200 pg/mL DL)*ELISA (17,000 pg/mL DL)
* The limit of detection (LOD) was calculated using 2.5 standard deviations from the 0 pg/mL calibrator.
** The standards were diluted in RPMI with 10% FBS.
Analyte Description SI2400 SI6000CHO Host Cell Protein Kit CHO Host Cell Protein Kit (1 Plate) K150HNF-1 K110HNF-1 CHO Host Cell Protein Kit (5 plates) K150HNF-2 K110HNF-2 CHO Host Cell Protein Kit (20 plates) K150HNF-3 K110HNF-3 CHO Host Cell Protein Base Kit (5 plates) K150HNA-3 K110HNA-3
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Kits and Assays for Host Cell Protein (HCP) Contamination
HCP Contamination Assay for Process Specific Applications (Customer Antibodies)
HCP Contamination Assay
Sign
al
Concentration (ng/mL)
10,000
100,000
1,000,000
0.1 1 10 100 1,000
WashedNo-Wash
10,0001,000
Biotin
Streptavidin
Host cellprotein
SULFO-TAG-labeledantibody
ElectrodeSurface
Biotin-labeledantibody
MSD users can develop contamination assays that incorporate their process-specific antibodies. Antibodies can be coated directly on MSD plates or immobilized through common biological linkers (e.g. streptavidin-biotin, anti-species antibodies). The HCP assay shown here was developed using customer antibodies in a bridging format. The assay can also be run in a no-wash format with only a modest (2.5 fold) loss in sensitivity.
No-Wash Assays for Routine Bioprocess Operation
Example Protocol 1. Add 25 µL/well of biotinylated anti-host cell protein (HCP)
antibody to MSD standard streptavidin plate.
2. Add 25 µL/well of SULFO-TAG™ labeled anti-HCP antibody, and add 25 µL/well of sample. Incubate for 2 hours.
3. Add 75 µL/well of 1X Read Buffer T and analyze plate on SECTOR instrument.
No-Wash HCP Contamination Assay
MSD (no-wash)
ELISA
10
200
7,500
5,000
LOD (ng/mL) Top Standard (ng/mL)
Sign
al
Concentration (ng/mL)
10,000
100,000
1,000,000
0.1 1 10 100 1,000 10,0001,000
MSD (wash)
MSD (no-wash)
3.9
10
7,500
7,500
LOD (ng/mL) Top Standard (ng/mL)
In routine operation, Bioprocess assays must be robust and simple. MSD assays have the sensitivity and reproducibility to exceed the require-ments of the applications, providing a margin of safety. Simple assay formats and wide dynamic ranges reduce time, labor and error by eliminat-ing dilutions and reducing wash steps.
This example shows the MSD CHO HCP contamination assay with a comparison to an ELISA using the same antibodies. It uses a bridging format where the same polyclonal anti-HCP antibody is used as both capture antibody (in biotinylated form) and detection antibody (SULFO-TAG labeled). The assay is 20 times more sensitive than the comparable ELISA. It also has a larger dynamic range
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Methotrexate is used to select high-producing cells. Although MTX helps generate higher yields, MTX contamination is a serious safety issue. MTX is often measured by HPLC, which is time consuming and requires concentration of samples to overcome limitations in sensitivity. MSD offers a method to replace HPLC assays. Our competitive immunoassay shows significantly improved sensitivity (0.01 ng/mL) compared to an HPLC method (1 ng/mL). The assay can be run in a one-wash or no-washed format. This assay incorporates a FITC-labeled MTX as the tracer and SULFO-TAG labeled anti-FITC as the detection antibody. This format is applicable to other competitive immunoassays.
Methotrexate (MTX) Contamination Kit
Kits and Assays for Impurities and Residual Contamination
Protein A can leach off columns during product purification. It is often challenging to detect since it may be bound to IgG proteins present in the sample. The MSD Protein A assay has a range from 50 pg/mL to 10 ng/mL when the sample matrix is free of IgG proteins. To buffer the effects of bound IgG, gamma globulin can be added to the assay diluents to extend the assay range from 500 pg/mL to 250 ng/mL. Dissociation protocols may make the assay more sensitive.
Protein A Contamination Kit
MSD Protein A Kit
Concentration [ng/mL]
Sign
al
0.0001 0.001 0.01 0.1 1 10 100 1,000
100
1,000
10,000
100,000
1,000,000
γ Globulin AddedNo lgG
MTX
Sign
al
Concentration [ng/mL]
0.001 0.001 0.001 0.001 0.0011,000
10,000
100,000
Free MTXSULFO-TAG-labeledanti-label antibody
Electrode surface
Labeled MTX
Directly immobilizedanti-MTX antibody
MSD Protocol 1. Add 20 µL of sample. Add 20 µL of FITC-labeled MTX. Incubate 2 hrs. 2. Add 25 µL of SULFO-TAG labeled anti-FITC. Incubate 2 hrs. 3. Wash plate 3x with PBS-T.
4. Add 150 µL of Read Buffer. Read.
Competitive Format
SULFO-TAG-labeledanti-Protein A antibody
Electrode surface
Protein A
Anti-Protein A antibody
MSD Protocol 1. Add 20 µL of sample. Incubate 2 hrs. 2. Add 25 µL of SULFO-TAG labeled anti-Protein A. Incubate 2hrs. 3. Wash plate 3x with PBS-T.
4. Add 150 µL of Read Buffer. Read.
Sandwich Format
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Multiplex Kits and Assays for Contamination
Residual Contamination Multiplex Kit
Protocol
1. Add 25 µL/well of standards (insulin and CHO-HCP) + 2 ng/mL of MTX-FITC tracer.
Incubate for 2 hours.
2. Add 25 µL/well of detection antibodies. Incubate for 2 hours.
3. Wash 3 times with PBS-T.
4. Add 150 µL/well of 1X Read Buffer T and analyze plate on SECTOR instrument.
Note: This assay was developed as a custom application. Performance of HCP contamination assays depends on the polyclonal antibodies used in the assay. Similiar assays can be developed using a variety of MSD products including the assay development packs listed on the back cover.
CHO-HCP Insulin
MTX
SULFO-TAG-anti-FITC
FITC-MTX
Anti-MTX
ECL
Coun
ts
1,000
10,000
1 100 10,000
MSD offers other residual protein assays such as insulin, protein A, methotrexate (MTX), cytokines, and extracellular matrix proteins. These assays are available in a single analyte and multiplex kits. Multiplexing combines many assays into a single plate without compromising performance, which leads to savings of both time and cost.
Concentration [pg/mL]
ECL
Coun
ts
1,000
10,000
10 1,000 100,000
Concentration [pg/mL]
ECL
Coun
ts
1,000
10,000
1 100 10,000
Concentration [pg/mL]
CHO Contamination Methotrexate Insulin Contamination
100,000
Detection Limit 300 pg/mL Detection Limit 20 pg/mL Detection Limit 40 pg/mL
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Assays for Antibody Screening and Production
MSD offers a suite of assays for antibody screening and production. Save time, cost, and sample by eliminating serial dilutions with MSD’s expanded dynamic range. Use multiplexing to improve early selection by combining measurements of abundance and specificity.
Protein A Coated Plates for IgG Measurements (Sandwich Assay)
SULFO-TAG-labeled F(ab)
lgG
Protein A plateProtein A Sandwich Assay
Concentration [ng/mL]
Sign
al
10,000
0.1 10 1,000
Concentration(ng/mL)
0
0.2
0.6
2.4
9.8
39
156
625
2,500
10,000
40,000
Average
672
686
698
808
1,165
2,581
8,906
46,835
146,705
200,520
223,810
Protein A Sandwich Assay
StdDev
15
17
9
10
45
121
283
2,634
4,494
13,416
6,817
%CV
2
3
1
1
4
5
3
6
3
7
3
100,000
1,000
Range of Assay 5 ng/mL - 10 µg/mL
LIGHTLIGHT
MSD Protocol
1. Add 25 µL of human IgG standard or sample. Incubate for 2
hours.
2. Wash 3x with PBS-T.
3. Add 25 µL of F(ab) anti-human detection antibody. Incubate
for 1 hour.
4. Wash 3x with PBS-T.
5. Add 150 µL of Read Buffer T. Read.
For increased sensitivity, MSD offers a sandwich immunoassay that uses a plate coated with Protein A (as the capture species) and SULFO-TAG labeled F(ab) goat-anti-human antibody as the detection species. The range of this assay is from 5 ng/mL to 10 µg/mL. This format can also be used to measure high levels of antibody with appropriate dilution.
Protein A Competition Assay
Concentration(µg/mL)
0
0.01
0.06
0.39
2.31
13.89
83.33
500
Average
15,489
15,156
12,829
9,900
4,160
1,753
733
362
Protein A Competition Assay
%CV
6.2
3.3
3.1
3.1
6.9
1.2
0.8
2.1
Detection Limit 0.04 µg/mL
Competition Protocol 1. Add 25 µL of sample. Add 25 µL of SULFO-TAG labeled
human IgG. Incubate 2 hours.
2. Add 100 µL of Read Buffer T (1.5x). Read.
SULFO-TAGlabeled
human IgG
Product
Protein A
Low Protein A
High Protein A
Concentration [µg/mL]
Sign
al
0.10 10 1,000
1,000
100
10,000
100,000
1,000,000 Protein A Coated
Tracer Concentration
2.1 ng
0.31 µg/mL
Protein A Coated Plates for IgG Measurements (Competition Assay)Our competitive immunoassays are designed for screening for high expressing clones and cells, where antibody levels are high (several mg/mL). MSD’s competitive assay uses a plate coated with Protein A to capture antibodies present in the sample and a SULFO-TAG-labeled human IgG as a tracer. The standard assay is completely homogeneous with a large dynamic range (from 0.5 µg/mL to 500 µg/mL). A modified version of the assay (which increases the amount of Protein A on the plate and raises the concentration of the tracer) can extend the upper limits of the dynamic range beyond 500 µg/mL (up to low mgs/mL).
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Assays for Antibody Screening and Production
lgG1
lgG2
lgG3
lgG4
lgA
lgM
Multiplex Isotyping Assay
lgG1 Spot
lg Concentration [pg/mL]
Sign
al 10,000
1,000 10,000
100,000
1,000
100,000
lgG2 Spot
lg Concentration [pg/mL]
Sign
al 1,000
1,000 10,000
10,000
100
100,000
lgG3 Spot
lg Concentration [pg/mL]
Sign
al
10,000
1,000 10,000
1,000
100,000
lgG4 Spot
lg Concentration [pg/mL]
Sign
al
10,000
1,000 10,000
1,000
100,000
lgA Spot
lg Concentration [pg/mL]
Sign
al 1,000
1,000 10,000
10,000
100
100,000
lgM Spot
lg Concentration [pg/mL]
Sign
al
10,000
1,000 10,000
1,000
100,000
Crossreactivity
Spot
lgG1
lgG2
lgG3
lgG4
lgA
lgM
100%
2.4%
0.6%
0.3%
0.0%
0.0%
0.3%
100%
0.5%
0.6%
1.1%
0.0%
lgG1 lgG2
0.4%
0.8%
100%
1.2%
0.1%
0.0%
lgG3
2.8%
2.7%
1.4%
100%
0.4%
0.0%
lgG4
0.1%
1.9%
0.5%
0.2%
100%
0.4%
lgA
Analyte
2.7%
2.7%
1.4%
0.9%
1.2%
100%
lgM
Heavy Chain - Light Chain Assay (λ Capture)
Concentration [µg/mL]
Sign
al
1001010.100.010
1,000,000
100,000
10,000
1,000
100
lgGlgG1lgG2lgG3lgG4
0
0.0021
0.0129
0.08
0.46
3
17
100
685.5
837
1,450.5
6,986
63,359
304,839
543,845
686,323
hIgG (mix)
662.5
656.5
662
709
768
1,389
9,581
102,471
hIgG1
664.5
673
752.5
736
1,003
2,453
11,041
70,397
hIgG2
670
1,150
3,741.5
22,752
156,329
486,752
730,099
858,279
hIgG3
693.5
908
2,582.5
27,159
214,939
504,625
613,057
652,288
hIgG4
AnalytehIgGµg/mL
Anti-Fc
Product
Anti-λ
LIGHT
lgG1
lgG2
lgG3
lgG4
lgA
lgM
lgG1
lgG2
lgG3
lgG4
lgA
lgM
lgG1
lgG2
lgG3
lgG4
lgA
lgM
lgG1
lgG2
lgG3
lgG4
lgA
lgM
lgG1
lgG2
lgG3
lgG4
lgA
lgM
Light Chain and Heavy Chain Immunoassays
MSD offers assays to detect intact (whole) antibodies. One format uses anti-λ or anti-κ light chain antibodies as capture antibodies. This assay detects intact antibodies with a large dynamic range and good sensitivity. The example below shows an assay with anti-λ capture antibody and a SULFO-TAG labeled anti-human heavy chain antibody. Purified human immunoglobulins were run along with a sample containing a cocktail of human immunoglobulins.
MSD’s custom multiplex assays for isotyping human antibodies have a large dynamic range that allows for measurement of isotypes in serum and plasma with simple dilution factors. The assays shown below use isotype-specific capture antibodies with a detection antibody. Purified isotypes were run on the panel in single concentrations to measure the cross-reactivity of the assays. The cross-reactivities were generally below 2% with the exception of IgG4 and IgM, which cross-reacted with IgG1 and IgG2 at about 3%. The specificity of the assay is difficult to measure because the purified standards may have levels of other isotypes present.
lgG1
lgG2
lgG3
lgG4
lgA
lgM
BSA
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Cell-Based Antibody Screening Assays
MSD enables screening of antibodies against cell surface proteins in whole cells or membranes.1 Cells readily bind to MSD plates through passive adsorption. These assays can be used in serology, hybridoma screening, to characterize relative binding affinities, to measure cell surface protein abundance, and for building functional binding assays for neutralizing antibodies. A variety of different cell lines, both adherent and suspension, have been tested including: CHO, HEK293, MCF7, BJAB, THP-1 and WIL. Assays can be run in 96-well or 384-well plates and can be run with 500-15,000 cells per well, offering a significant reduction in the number of cells used. The cells are not fixed or processed, so receptors maintain their natural conformation. Several assays have been developed with a single or no-wash format.
AB1AB2AB3
Sign
al (t
rans
fect
ed) -
Sig
nal (
wild
typ
e)
Concentration of Antibody (µg/mL)
5,000
7,000
0.2 0.4 0.6 0.8 10
4,000
3,000
2,000
1,000
6,000
1.2-1,000
Positive 1
Positive 2
Negative
0
SULFO-TAG-labeledanti-lgG
Antibody
Cells
Carbon electrode
Cell surface protein
MSD Protocol
1. Add 12,500 cells per well in 50 µL volume. Incubate for 2 hours.
2. Dump out plate. Add 25 µL of cell supernatant. Add 25 µL of SULFO-TAG labeled goat-anti-mouse. Incubate for 2 hours.
3. Wash plate 3x with PBS.
4. Add 150 µL of Read Buffer T (without surfactant). Read.
MSD assays can replace FACS analysis for screening antibodies against cell-surface receptors. Unlike FACS, MSD assays do not require purification of receptors, a significant advantage in method development. MSD assays also save direct costs with less expensive reagents.
MSD Assays to Replace FACS Methods
Mice were inoculated with whole cells to generate antibodies against the extracellular domain of a cell surface protein. The relative affinities of the two positives matched those found by FACS analysis. When compared to the FACS protocol, the MSD protocol used about 10 fold less cells with a 20 fold increase in throughput. An entire batch of screening required 1 week through FACS, but could be accomplished in one day on the MSD system.
1 Lu, Y., Wong, W.L., Meng, Y.G. (2006) A high throughput electrochemiluminescent cell-binding assay for therapeutic anti-CD20 antibody selection. Journal of Immunology Methods.
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Immunogenicity testing is a crucial part of biopharmaceutical development. More stringent recommendations regarding immunogenicity assay performance necessitates the development of more robust and tolerant assays. MSD assays exhibit excellent sensitivity, precision, free drug tolerance, and minimal matrix effects. In addition, MSD assays are capable of finding low affinity antibodies during initial screens, and have a large linear range that reduces the number of required sample dilutions. Build assays for many drug types using MSD technology, including antibodies, humanized antibodies, proteins, and peptides with reagents designed to provide a variety of flexible assay formats and facilitate rapid assay development. Comparisons of MSD immunogenicity assays to the traditional ELISA format are featured below, using both a bridging assay format and direct immobilization format. Visit www.mesoscale.com for more information on immunogenicity assay develop-ment and a complete listing of materials and reagents.
Direct Immunogenicity Assays for Protein Drugs
MSD Bridging Assay Protocol1. Combine biotin-drug, sTAG-drug and sample in polypropylene plate and
incubate 1 hour to overnight. 2. Transfer solution to pre-blocked standard streptavidin MSD plate. Incubate
for 1 hour. 3. Wash assay plate; add Read Buffer T; read plate on SECTOR instrument.
MSD Sandwich Immunogenicity Assay Protocol
1. Coat plate with drug at 0.05 to 5 pmole per well and incubate for 1 hour to overnight.
2. Block with 150 mL for 1 hour. 3. Wash plate. Add 25 mL of sample. 4. (Optional wash). Add 25 mL of detection antibody. 5. Wash assay plate; add Read Buffer T; read plate on SECTOR instrument.
Neat human serum was used as the sample matrix. The top of the curve is about1 µg/mL for both formats, but the MSD format is 40 times more sensitive.
Bridging Immunogenicity Assay: ELISA Comparison
Sign
al
ADA Concentration (ng/mL)
100,000
10,000
0.01 10 10,000
1,000
MSDELISA
1,000,000
Sign
al/B
ackg
roun
d
ADA Concentration [ng/ml]
MSD assay shows comparable sensitivity to ELISA, with a larger dynamic range and a simple homogenous incubation.
Reference: Moxness, M., Tatarewicz, S., Weeraratne, D., Murakami, N., Wullner, D., Mytych, D., Jawa, V., Koren, E., Swanson, S.J. (2005) Immunogenicity Testing by Electrochemiluminescent Detection for Antibodies Directed against Therapeutic Human Monoclonal Antibodies. Clinical Chemistry. 51: 1983-1985.
MSDELISA
1,000
100
10
1
0.1 10 1,000
SULFO-TAGlabeled drug
Anti-drugantibody
Biotin-labeleddrug
MSD SA plate
SULFO-TAGanti-human
antibody
Anti-drugantibody
Protein drug
Better Free Drug Tolerance
Detection of Low Affinity Antibodies
Fewer Washes
High-Throughput
Direct Conjugation of Stable Label
Improved Sensitivity
Increased Dynamic Range
Reduced Sample Volume
Higher Binding Capacity
Poor
No
3-4
Good
Yes
100s ng/mL
1-2 logs
25-100 µL
Excellent
Yes
1
High
Yes
10s ng/mL
3-4 logs
5-25 µL
10X More
ELISA MSD
Better Free Drug Tolerance
Detection of Low Affinity Antibodies
Fewer Washes
High-Throughput
Direct Conjugation of Stable Label
Improved Sensitivity
Increased Dynamic Range
Reduction in Reagent Consumption
Higher Binding Capacity
Poor
No
3-5
Good
No
100s ng/mL
1-2 logs
Good
Maybe
2-3
Good
No
10s ng/mL
3-4.5 logs
2-10 fold
10X More
ELISA MSD
Immunogenicity
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Functional Cell Based Assays and Assays for Potency
Cytokines
IL-12p70IL-13IL-8TNF-αIL-10
IL-4IFN-γ
IL-5IL-1β
IL-2Capture Ab
Electrode
IL-8
Labeled-Ab
MSD Human TH1/TH2
Colormap display demonstrates different patterns of cytokine responses in different wells of a 96-well 10-spot assay plate. 10 - 10,000 counts
MSD cytokine assays are available in single- and multi-analyte kits. MSD’s cytokine protocols are designed to optimize workflow and ease-of-use while maximizing assay performance. These protocols have been used successfully for many clinical and animal sample matrices including cell culture supernatants, serum, plasma, sputum, bronchoalveolar lavage, and other bodily fluids.
TNF-α
0.1
0.1
IL-10
0.5
0.5
IL-12p70
0.5
0.7
IL-13
0.6
0.3
IL-4
0.3
0.6
IL-5
0.2
0.1
IL-8
0.4
1.2
IFN-γ
0.5
0.9
IL-1β
0.3
0.3
IL-2
0.2
0.2
Detection Parameters (pg/mL)
LLOD
LLOQ
IFN-γIL-1βIL-2IL-4IL-5IL-8IL-10IL-12p70IL-13TNF-α
Sign
alConcentration (pg/mL)
1,000,000
100,000
10,000
1,000
100
0.01 1 10,000100
Ultra-Sensitive Kit10,000,000
pVEGFR-2
Traditional Western
20 μg lysate per lane
Pos
VEGFR-2
Neg
MSD Experimental
1.3 μg lysate per well
Pos Neg
0 0.15 1.3 50
15,000
30,000
Mea
n Si
gnal
Lysates [μg]102.50.30.08
20,000
10,000
25,000
5,000
0.6
VEGFR-2 (pTyr1054/1059)Positive
Negative
Phosphoproteins and Biomarkers
SULFO-TAG antibody
Analyte
Capture antibody
Cell based bioassays often use proliferation or apoptosis to measure the functional response to a drug product. These assays can be extremely variable and dependent on the passage of cell line and media. An alternative methodology is to measure receptor phosphorylation or secreted biomarkers. Receptor phosphorylation assay can be performed by stimulating the cells within a plate with the drug, lysing the cells, and detecting the phosphorylation event with an MSD kit. Secreted biomarkers can be measured in complex matrices such as serum and plasma.
Sign
al
Concentration [pg/mL]
bFGF
101
104
102
106
10-2 10-1 100 102 104103101
103
105
Sign
al
Concentration [pg/mL]
VEGF
102
103
105
10-2 10-1 100 102 104103101
104
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Catalog Numbers
Visit www.mesoscale.com for a complete product listing.
Analyte Description SI2400 SI6000Protein A Protein A Contamination Assay Kit (1 Plate) K150HMF-1 K110HMF-1 Protein A Contamination Assay Kit (5 plates) K150HMF-2 K110HMF-2 Protein A Contamination Assay Kit (20 plates) K150HMF-3 K110HMF-3 Protein A Contamination Assay Base Kit (5 plates) K150HMA-3 K110HMA-3 CHO Host Cell Protein CHO Host Cell Protein Kit (1 Plate) K150HNF-1 K110HNF-1 CHO Host Cell Protein Kit (5 Plates) K150HNF-2 K110HNF-2 CHO Host Cell Protein Kit (20 Plates) K150HNF-3 K110HNF-3 CHO Host Cell Protein Base Kit (20 Plates) K150HNA-3 K110HNA-3 Methotrexate Methotrexate Kit (1 Plate) K150H0F-1 K110H0F-1 Methotrexate Kit (5 Plates) K150H0F-2 K110H0F-2 Methotrexate Kit (20 Plates) K150H0F-3 K110H0F-3 Methotrexate Base Kit (20 Plates) K150H0A-3 K110H0A-3 Protein A, CHO Host Cell Protein BioProcess Duplex Kit (1 Plate) K15149F-1 K11149F-1 BioProcess Duplex Kit (5 Plates) K15149F-2 K11149F-2 BioProcess Duplex Kit (20 Plates) K15149F-3 K11149F-3 BioProcess Duplex Base Kit (20 Plates) K15149A-3 K11149A-3 Insulin, Methotrexate, CHO Host Cell Protein BioProcess 3-plex Kit (1 Plate) K15150F-1 K11150F-1 BioProcess 3-plex Kit (5 Plates) K15150F-2 K11150F-2 BioProcess 3-plex Kit (20 Plates) K15150F-3 K11150F-3 BioProcess 3-plex Base Kit (20 Plates) K15150A-3 K11150A-3
Bioprocess Assays
ELISA ConversionPack I (Uncoated Plates)
ELISA ConversionPack II (Anti-species Plates)
ELISA ConversionPack III (Avidin/Streptavidin Plates)
Immunogenicity Development Pack
Cell Based Assays Development Pack
K13A01-1
K13A02-1
K13A03-1
K13A04-1
K13A05-1
K17A01-1
K17A02-1
K17A03-1
K17A04-1
K17A05-1
K15A01-1
K15A02-1
K15A03-1
K15A04-1
K15A05-1
K11A01-1
K11A02-1
K11A03-1
K11A04-1
K11A05-1
Immunoassay Development Kits (96-well Plates)
Description PR100 PR400 SI2400 SI6000
Purchase orders should be faxed or emailed to the following queries: TEL: 1.240.631.2522 x4685FAX: 1.301.990.2776EMAIL: [email protected]
Contact Information
13
Catalog Numbers
Coating Plate Format Binding Capacity Description PR100 PR400
96-well Plate L13XA-1 L17XA-196-well Small Spot Plate L43XA-1 L47XA-196-well High-Bind Plate L13XB-1 L17XB-196-well High-Bind Small Spot Plate L43XB-1 L47XB-1
96-well Avidin Plate L13AA-1 L17AA-196-well High-Bind Avidin Plate L13AB-1 L17AB-1
96-well Streptavidin Plate L13SA-1 L17SA-196-well High-Bind Streptavidin Plate L13SB-1 L17SB-1
96-well High-Bind Glutathione Plate L13GB-1 L17GB-1
96-well GAR Plate L13RA-1 L17RA-1
96-well GAM Plate L13MA-1 L17MA-1
9696 SS9696 SS
9696
9696
96
96
96
StandardStandardHigh BindHigh Bind
StandardHigh Bind
StandardHigh Bind
High Bind
Standard
Standard
Bare
Avidin
Streptavidin
Glutathione
Anti-Rabbit
Anti-Mouse
MULTI-ARRAY Plates for the SECTOR PR Readers
Coating Plate Format Binding Capacity Description SI2400 SI6000
96-well Plate L15XA-1 L11XA-196-well Small Spot Plate L45XA-1 L41XA-1384-well Plate L25XA-1 L21XA-196-well High-Bind Plate L15XB-1 L11XB-196-well High-Bind Small Spot Plate L45XB-1 L41XB-1384-well High-Bind Plate L25XB-1 L21XB-1
96-well Avidin Plate L15AA-1 L11AA-1384-well Avidin Plate L25AA-1 L21AA-196-well High-Bind Avidin Plate L15AB-1 L11AB-1384-well High-Bind Avidin Plate L25AB-1 L21AB-1
96-well Streptavidin Plate L15SA-1 L11SA-1384-well Streptavidin Plate L25SA-1 L21SA-196-well High-Bind Streptavidin Plate L15SB-1 L11SB-1384-well High-Bind Streptavidin Plate L25SB-1 L21SB-1
96-well High-Bind Glutathione Plate L15GB-1 L11GB-1384-well High-Bind Glutathione Plate L25GB-1 L21GB-1
96-well Small Spot GAR Plate L45RA-1 L41RA-1384-well GAR Plate L25RA-1 L21RA-1
96-well Small Spot GAM Plate L45MA-1 L41MA-1384-well GAM Plate L25MA-1 L21MA-1
9696 SS3849696 SS384
9638496384
9638496384
96384
96 SS384
96 SS384
StandardStandardStandardHigh BindHigh BindHigh Bind
StandardStandardHigh BindHigh Bind
StandardStandardHigh BindHigh Bind
High BindHigh Bind
StandardStandard
StandardStandard
Bare
Avidin
Streptavidin
Glutathione
Anti-Rabbit(Goat)
Anti-Mouse(Goat)
MULTI-ARRAY Plates for the SECTOR Imagers
Protein A 96-well Protein A Plate L13DB-1 L17DB-196 High Bind
96-well Protein A Plate L15DB-1 L11DB-1384-well Protein A Plate L25DB-1 L21DB-1
96 384
High BindHigh Bind
Protein A
14
Catalog Numbers
Ruthenium Products
MSD Label Reporter Quantity Catalog Number
50 µg200 µg
50 µg200 µg
50 µg200 µg
50 µg200 µg
200 µg
200 µg
500 µg
R32AB-5R32AB-1
R32AC-5R32AC-1
R32AJ-5R32AJ-1
R32AD-5R32AD-1
R31AB-1
R31AC-1
R31AD-2
Anti-Rabbit Antibody (Goat)Anti-Rabbit Antibody (Goat)
Anti-Mouse Antibody (Goat)Anti-Mouse Antibody (Goat)
Anti-Human Antibody (Goat)Anti-Human Antibody (Goat)
StreptavidinStreptavidin
Anti-Rabbit Antibody (Goat)
Anti-Mouse Antibody (Goat)
Streptavidin
SULFO-TAGSULFO-TAG
SULFO-TAGSULFO-TAG
SULFO-TAGSULFO-TAG
SULFO-TAGSULFO-TAG
TAG
TAG
TAG
Description Quantity Catalog Number
R91AN-1R91AN-2R91BN-1R91BN-2
150 nMoles500 nMoles150 nMoles500 nMoles
SULFO-TAGSULFO-TAGTAGTAG
Conjugated Reporters Labeling Reagents (NHS Ester)
Description Surfactant Quantity Catalog Number
50 mL200 mL1 L
50 mL200 mL1 L
50 mL200 mL1 L
50 mL200 mL1 L
50 mL200 mL1 L
50 mL200 mL1 L
50 mL200 mL1 L
R92TC-3R92TC-2R92TC-1
R92TD-3R92TD-2R92TD-1
R92PC-3R92PC-2R92PC-1
R92SC-3R92SC-2R92SC-1
R92SD-3R92SD-2R92SD-1
R92GC-3R92GC-2R92GC-1
R92GD-3R92GD-2R92GD-1
+++
---
+++
+++
---
+++
---
Read Buffer T (4X)Read Buffer T (4X)Read Buffer T (4X)
Read Buffer T (4X)Read Buffer T (4X)Read Buffer T (4X)
Read Buffer P (4X)Read Buffer P (4X)Read Buffer P (4X)
Read Buffer S (4X)Read Buffer S (4X)Read Buffer S (4X)
Read Buffer S (4X)Read Buffer S (4X)Read Buffer S (4X)
Read Buffer G (4X)Read Buffer G (4X)Read Buffer G (4X)
Read Buffer G (4X)Read Buffer G (4X)Read Buffer G (4X)
Description Quantity Catalog #
250 mL1 L
50 mL200 mL1 L
50 mL200 mL
200 mL1 L
R93AA-2R93AA-1
R50AA-4R50AA-2R50AA-3
R60TX-3R60TX-2
R61TX-2R61TX-1
Blocker A Kit
Antibody Diluent
Tris Lysis Buffer
Tris Wash Buffer (10X)
Read Buffers Diluents and Buffers
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For research use only; not for use in diagnostic or therapeutic procedures.
®
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