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

2

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

3

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

4

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

5

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

6

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).

8

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

9

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.

10

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

11

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

12

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: customerservice@mesoscale.com

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

+++

---

+++

+++

---

+++

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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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