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. MSDs 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 Discovery

A division of Meso Scale Diagnostics, LLC.
9238 Gaither Road Gaithersburg, MD USA 20877
Phone: 240.631.2522 (x4685 for customer service)
www.mesoscale.com
MSD Technology

MSDs 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

MULTI-ARRAY Plate
B

LIGHT Measured signal is light

Luminescence
Electrochemiluminescence Features: *Ru(bpy)32+
Emitting Light

-H+
TPA.
Minimal background signals and high signal to back- Chemi-
Chemical Energy
ground ratios - the stimulation mechanism (electricity) is Ru(bpy)32+
Ru(bpy)3 3+ TPA TPA.+
decoupled from the signal (light)
Electro-
Electrochemically Initiated
e- e-

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 Dielectric

Counter Electrode Working Electrode
quenching

Signal amplification - multiple excitation cycles of each

label enhance light levels and improve sensitivity

MULTI-ARRAY and MULTI-SPOT Features:

Flexible surface coatings to suit most any biology High density arrays for high content screening and high throughput
multiplexing of biomarkers
Carbon electrode plate surface has 10X greater binding
capacity than polystyrene Custom surface coatings and patterns

MULTI-SPOT Assay
Analyte 1
Simultaneous measure-
ment of analytes in one
Dielectric Analyte 2
well. Capture antibodies
are arrayed on the
patterned working Counter electrode Analyte 3
electrode. Analytes are
detected with antibodies Analyte 4
labeled with SULFO-TAG
reagent.
2
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.

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.

MSD vs ELISA-CHO HCP Assay

10,000
0.8
Absorbance (492 nm-405 nm)

MSD (200 pg/mL DL)*

ELISA (17,000 pg/mL DL) SULFO-TAG-
0.65 labeled anti-CHO
HCP antibody
0.55
Signal

Host cell
0.45 protein

0.35 Anti-CHO HCP

0.3
capture antibody
1,000

0.25
Electrode Surface
10 100 1,000 10,000 100,000

Concentration (pg/mL)**

* 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.

CHO Host Cell Protein Kit (96-well plates)

Analyte Description SI2400 SI6000
CHO 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

3
Kits and Assays for Host Cell Protein (HCP) Contamination

HCP Contamination Assay for Process Specific Applications (Customer Antibodies)

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.

HCP Contamination Assay

1,000,000 SULFO-TAG-
labeled
Washed antibody
No-Wash

Host cell
100,000 protein

Biotin-labeled
Signal

antibody

Biotin
10,000
Streptavidin
Electrode
Surface

1,000
LOD (ng/mL) Top Standard (ng/mL)
0.1 1 10 100 1,000 10,000
MSD (wash) 3.9 7,500
Concentration (ng/mL)
MSD (no-wash) 10 7,500

No-Wash Assays for Routine Bioprocess Operation

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

No-Wash HCP Contamination Assay

1,000,000 LOD (ng/mL) Top Standard (ng/mL)
MSD (no-wash) 10 7,500
ELISA 200 5,000

100,000
Example Protocol
Signal

1. Add 25 L/well of biotinylated anti-host cell protein (HCP)

antibody to MSD standard streptavidin plate.
10,000
2. Add 25 L/well of SULFO-TAG labeled anti-HCP antibody, and
add 25 L/well of sample. Incubate for 2 hours.

3. Add 75 L/well of 1X Read Buffer T and analyze plate on SECTOR

1,000 instrument.

0.1 1 10 100 1,000 10,000

Concentration (ng/mL)

4
Kits and Assays for Impurities and Residual Contamination

Methotrexate (MTX) Contamination Kit

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.

MTX Competitive Format

100,000

SULFO-TAG-labeled
anti-label antibody Free MTX

Labeled MTX
Signal

10,000 Directly immobilized

anti-MTX antibody

Electrode surface

MSD Protocol
1,000
1. Add 20 L of sample. Add 20 L of FITC-labeled MTX. Incubate 2 hrs.
0.001 0.001 0.001 0.001 0.001
Concentration [ng/mL] 2. Add 25 L of SULFO-TAG labeled anti-FITC. Incubate 2 hrs.

3. Wash plate 3x with PBS-T.

4. Add 150 L of Read Buffer. Read.

Protein A Contamination Kit

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.

MSD Protein A Kit Sandwich Format

1,000,000
Globulin Added SULFO-TAG-labeled
No lgG anti-Protein A antibody
100,000
Protein A
Signal

10,000 Anti-Protein A antibody

Electrode surface
1,000

MSD Protocol
100
1. Add 20 L of sample. Incubate 2 hrs.
0.0001 0.001 0.01 0.1 1 10 100 1,000
Concentration [ng/mL] 2. Add 25 L of SULFO-TAG labeled anti-Protein A. Incubate 2hrs.

3. Wash plate 3x with PBS-T.

4. Add 150 L of Read Buffer. Read.

5
 Multiplex Kits and Assays for Contamination

Residual Contamination Multiplex Kit

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.

CHO-HCP Insulin

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
MTX developed using a variety of MSD products including the assay
development packs listed on the back cover.

Protocol
1. Add 25 L/well of standards (insulin and CHO-HCP) + 2 ng/mL of MTX-FITC tracer.
Incubate for 2 hours.
SULFO-TAG-
anti-FITC 2. Add 25 L/well of detection antibodies. Incubate for 2 hours.

3. Wash 3 times with PBS-T.

FITC-MTX 4. Add 150 L/well of 1X Read Buffer T and analyze plate on SECTOR instrument.

Anti-MTX

CHO Contamination Methotrexate Insulin Contamination

100,000
10,000
10,000
ECL Counts

ECL Counts
ECL Counts

10,000

1,000
1,000
1,000

1 100 10,000 10 1,000 100,000 1 100 10,000

Concentration [pg/mL] Concentration [pg/mL] Concentration [pg/mL]

Detection Limit 300 pg/mL Detection Limit 20 pg/mL Detection Limit 40 pg/mL

6
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 MSDs
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) LIGHT

LIGHT

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 SULFO-TAG-
the detection species. The range of this assay is from 5 ng/mL to 10 g/mL. This format can labeled F(ab)
also be used to measure high levels of antibody with appropriate dilution. lgG

Protein A Sandwich Assay

Protein A plate
Concentration Protein A Sandwich Assay
(ng/mL) Average StdDev %CV
100,000
0 672 15 2 MSD Protocol
0.2 686 17 3
Signal

0.6 698 9 1 1. Add 25 L of human IgG standard or sample. Incubate for 2

10,000 2.4 808 10 1
hours.

9.8 1,165 45 4
2. Wash 3x with PBS-T.
39 2,581 121 5
156 8,906 283 3
1,000 3. Add 25 L of F(ab) anti-human detection antibody. Incubate
625 46,835 2,634 6
for 1 hour.
2,500 146,705 4,494 3
0.1 10 1,000 10,000 200,520 13,416 7
Concentration [ng/mL] 40,000 223,810 6,817 3
4. Wash 3x with PBS-T.

5. Add 150 L of Read Buffer T. Read.

Range of Assay 5 ng/mL - 10 g/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 SULFO-TAG
labeled
mg/mL). MSDs competitive assay uses a plate coated with Protein A to human IgG
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 g/mL to 500 g/mL). A modified Product
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 Protein A
the dynamic range beyond 500 g/mL (up to low mgs/mL).

Protein A Competition Assay

1,000,000 Protein A Coated 2.1 ng
Low Protein A Concentration Protein A Competition Assay
(g/mL) Average %CV Tracer Concentration 0.31 g/mL
High Protein A
100,000 0 15,489 6.2
0.01 15,156 3.3
Competition Protocol
Signal

0.06 12,829 3.1

10,000 0.39 9,900 3.1 1. Add 25 L of sample. Add 25 L of SULFO-TAG labeled
2.31 4,160 6.9 human IgG. Incubate 2 hours.
1,000 13.89 1,753 1.2
83.33 733 0.8 2. Add 100 L of Read Buffer T (1.5x). Read.
500 362 2.1
100
0 0.1 10 1,000
Detection Limit 0.04 g/mL
Concentration [g/mL]

7
Assays for Antibody Screening and Production

Light Chain and Heavy Chain Immunoassays LIGHT

MSD offers assays to detect intact (whole) antibodies. One format uses anti- or anti- 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- capture antibody and a SULFO-TAG labeled anti-human heavy Product
chain antibody. Purified human immunoglobulins were run along with a sample containing a cocktail of Anti-Fc
Anti-
human immunoglobulins.

Heavy Chain - Light Chain Assay ( Capture)

1,000,000 hIgG Analyte

lgG g/mL
hIgG (mix) hIgG1 hIgG2 hIgG3 hIgG4
lgG1
0 685.5 662.5 664.5 670 693.5
100,000 lgG2
lgG3 0.0021 837 656.5 673 1,150 908
lgG4
Signal

0.0129 1,450.5 662 752.5 3,741.5 2,582.5

10,000 0.08 6,986 709 736 22,752 27,159
0.46 63,359 768 1,003 156,329 214,939
3 304,839 1,389 2,453 486,752 504,625
1,000
17 543,845 9,581 11,041 730,099 613,057
100 686,323 102,471 70,397 858,279 652,288
100
0 0.01 0.10 1 10 100
Concentration [g/mL]

Multiplex Isotyping Assay

MSDs 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 Spot lgG2 Spot lgG3 Spot Crossreactivity

10,000
100,000 lgG1 lgG1 lgG1 Analyte
lgG2 lgG2 lgG2
lgG3 lgG3 10,000 lgG3 Spot lgG1 lgG2 lgG3 lgG4 lgA lgM
lgG4 lgG4 lgG4
lgG1
Signal

100% 0.3%
Signal

Signal

10,000 lgA
1,000
lgA lgA 0.4% 2.8% 0.1% 2.7%
lgM lgM lgM
lgG2 2.4% 100% 0.8% 2.7% 1.9% 2.7%
1,000 lgG3 0.6% 0.5% 100% 1.4% 0.5% 1.4%
1,000
lgG4 0.3% 0.6% 1.2% 100% 0.2% 0.9%
100
lgA 0.0% 1.1% 0.1% 0.4% 100% 1.2%
1,000 10,000 100,000 1,000 10,000 100,000 1,000 10,000 100,000
lgM 0.0% 0.0% 0.0% 0.0% 0.4% 100%
lg Concentration [pg/mL] lg Concentration [pg/mL] lg Concentration [pg/mL]

lgG4 Spot lgA Spot lgM Spot

10,000
lgG1 lgG1 lgG1
lgG2 lgG2 lgG2
lgG3 lgG3 10,000 lgG3 lgG1
10,000 lgG4 lgG2
lgG4 lgG4
lgG3
Signal
Signal

Signal

lgA lgA lgA

lgM
1,000 lgM lgM lgG4
lgA
1,000 1,000
lgM
BSA

100

1,000 10,000 100,000 1,000 10,000 100,000 1,000 10,000 100,000

lg Concentration [pg/mL] lg Concentration [pg/mL] lg Concentration [pg/mL]

8
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.

SULFO-TAG-labeled
anti-lgG

Antibody

Cell surface protein

Cells

Carbon electrode

MSD Assays to Replace FACS Methods

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.
Signal (transfected) - Signal (wild type)

7,000 Mice were inoculated with whole cells to generate antibodies

AB1 Positive 1
6,000 against the extracellular domain of a cell surface protein. The
AB2 relative affinities of the two positives matched those found by
5,000 AB3 FACS analysis. When compared to the FACS protocol, the
4,000 MSD protocol used about 10 fold less cells with a 20 fold
increase in throughput. An entire batch of screening required
3,000 1 week through FACS, but could be accomplished in one day
2,000 on the MSD system.
Positive 2
1,000
Negative MSD Protocol
0
1. Add 12,500 cells per well in 50 L volume. Incubate for 2 hours.
0 0.2 0.4 0.6 0.8 1 1.2
-1,000 2. Dump out plate. Add 25 L of cell supernatant. Add 25 L of
Concentration of Antibody (g/mL) SULFO-TAG labeled goat-anti-mouse. Incubate for 2 hours.
3. Wash plate 3x with PBS.
4. Add 150 L of Read Buffer T (without surfactant). Read.

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.

9
Immunogenicity

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.

ELISA MSD
Bridging Immunogenicity Assay: ELISA Comparison
Better Free Drug Tolerance Poor Excellent
1,000,000 Detection of Low Affinity Antibodies No Yes
MSD Fewer Washes 3-4 1
ELISA High-Throughput Good High
SULFO-TAG
100,000 labeled drug Direct Conjugation of Stable Label Yes Yes
Anti-drug Improved Sensitivity 100s ng/mL 10s ng/mL
Signal

antibody
Increased Dynamic Range 1-2 logs 3-4 logs
10,000
Biotin-labeled Reduced Sample Volume 25-100 L 5-25 L
drug
Higher Binding Capacity 10X More
1,000 MSD SA plate
MSD Bridging Assay Protocol
1. Combine biotin-drug, sTAG-drug and sample in polypropylene plate and
0.01 10 10,000 incubate 1 hour to overnight.
ADA Concentration (ng/mL)
2. Transfer solution to pre-blocked standard streptavidin MSD plate. Incubate
for 1 hour.
MSD assay shows comparable sensitivity to ELISA, with a larger dynamic range and a
simple homogenous incubation.
3. Wash assay plate; add Read Buffer T; read plate on SECTOR instrument.

Direct Immunogenicity Assays for Protein Drugs

ELISA MSD

1,000 Better Free Drug Tolerance Poor Good

MSD Detection of Low Affinity Antibodies No Maybe
ELISA
Fewer Washes 3-5 2-3
Signal/Background

100 High-Throughput Good Good

SULFO-TAG
anti-human Direct Conjugation of Stable Label No No
antibody
Improved Sensitivity 100s ng/mL 10s ng/mL
10 Anti-drug Increased Dynamic Range 1-2 logs 3-4.5 logs
antibody
Reduction in Reagent Consumption 2-10 fold
Protein drug Higher Binding Capacity 10X More
1
MSD Sandwich Immunogenicity Assay Protocol
0.1 10 1,000
ADA Concentration [ng/ml] 1. Coat plate with drug at 0.05 to 5 pmole per well and incubate for 1 hour
to overnight.
Neat human serum was used as the sample matrix. The top of the curve is about 2. Block with 150 mL for 1 hour.
1 g/mL for both formats, but the MSD format is 40 times more sensitive.
3. Wash plate. Add 25 mL of sample.

4. (Optional wash). Add 25 mL of detection antibody.

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 5. Wash assay plate; add Read Buffer T; read plate on SECTOR instrument.
Human Monoclonal Antibodies. Clinical Chemistry. 51: 1983-1985.

10
Functional Cell Based Assays and Assays for Potency

Cytokines
MSD cytokine assays are available in single- and multi-analyte kits. MSDs 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.

MSD Human TH1/TH2

Ultra-Sensitive Kit
IL-4 IL-12p70 Labeled-Ab
10,000,000
IFN- IL-13
IL-8
IL-5 IL-8 IFN-
IL-1 TNF- Capture Ab 1,000,000 IL-1
IL-2 IL-10 IL-2
Electrode
100,000 IL-4
IL-5

Signal
IL-8
10,000
IL-10
Colormap display demonstrates different patterns of cytokine
responses in different wells of a 96-well 10-spot assay plate. IL-12p70
10 - 10,000 counts 1,000 IL-13
TNF-
Detection Parameters (pg/mL) 100

IFN- IL-1 IL-2 IL-4 IL-5 IL-8 IL-10 IL-12p70 IL-13 TNF- 0.01 1 100 10,000
LLOD 0.5 0.3 0.2 0.3 0.2 0.4 0.5 0.5 0.6 0.1 Concentration (pg/mL)
LLOQ 0.9 0.3 0.2 0.6 0.1 1.2 0.5 0.7 0.3 0.1

Phosphoproteins and Biomarkers

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.

30,000 Positive VEGFR-2 (pTyr1054/1059)

25,000 Negative
Traditional Western MSD Experimental
Pos Neg
Pos Neg
Mean Signal

20,000
pVEGFR-2
15,000

10,000 VEGFR-2

5,000 20 g lysate per lane 1.3 g lysate per well

0
0 0.08 0.15 0.3 0.6 1.3 2.5 5 10
Lysates [g]

bFGF VEGF
106 105

105
SULFO-TAG antibody
104
104
Signal

Signal

Analyte
103
103
Capture antibody
102

101 102
10-2 10-1 100 101 102 103 104 10-2 10-1 100 101 102 103 104
Concentration [pg/mL] Concentration [pg/mL]

11
Catalog Numbers

Bioprocess Assays

Analyte Description SI2400 SI6000

Protein 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

Immunoassay Development Kits (96-well Plates)

Description PR100 PR400 SI2400 SI6000
ELISA Conversion K13A01-1 K17A01-1 K15A01-1 K11A01-1
Pack I (Uncoated Plates)

ELISA Conversion K13A02-1 K17A02-1 K15A02-1 K11A02-1

Pack II (Anti-species Plates)

ELISA Conversion K13A03-1 K17A03-1 K15A03-1 K11A03-1

Pack III (Avidin/Streptavidin Plates)

Immunogenicity Development Pack K13A04-1 K17A04-1 K15A04-1 K11A04-1

Cell Based Assays Development Pack K13A05-1 K17A05-1 K15A05-1 K11A05-1

Visit www.mesoscale.com for a complete product listing.

Contact Information
Purchase orders should be faxed or emailed to the following queries:

TEL: 1.240.631.2522 x4685

FAX: 1.301.990.2776
EMAIL: customerservice@mesoscale.com

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Catalog Numbers

MULTI-ARRAY Plates for the SECTOR PR Readers

Coating Plate Format Binding Capacity Description PR100 PR400
Bare 96 Standard 96-well Plate L13XA-1 L17XA-1
96 SS Standard 96-well Small Spot Plate L43XA-1 L47XA-1
96 High Bind 96-well High-Bind Plate L13XB-1 L17XB-1
96 SS High Bind 96-well High-Bind Small Spot Plate L43XB-1 L47XB-1

Avidin 96 Standard 96-well Avidin Plate L13AA-1 L17AA-1

96 High Bind 96-well High-Bind Avidin Plate L13AB-1 L17AB-1

Streptavidin 96 Standard 96-well Streptavidin Plate L13SA-1 L17SA-1

96 High Bind 96-well High-Bind Streptavidin Plate L13SB-1 L17SB-1

Glutathione 96 High Bind 96-well High-Bind Glutathione Plate L13GB-1 L17GB-1

Anti-Rabbit 96 Standard 96-well GAR Plate L13RA-1 L17RA-1

Anti-Mouse 96 Standard 96-well GAM Plate L13MA-1 L17MA-1

Protein A 96 High Bind 96-well Protein A Plate L13DB-1 L17DB-1

MULTI-ARRAY Plates for the SECTOR Imagers

Coating Plate Format Binding Capacity Description SI2400 SI6000
Bare 96 Standard 96-well Plate L15XA-1 L11XA-1
96 SS Standard 96-well Small Spot Plate L45XA-1 L41XA-1
384 Standard 384-well Plate L25XA-1 L21XA-1
96 High Bind 96-well High-Bind Plate L15XB-1 L11XB-1
96 SS High Bind 96-well High-Bind Small Spot Plate L45XB-1 L41XB-1
384 High Bind 384-well High-Bind Plate L25XB-1 L21XB-1

Avidin 96 Standard 96-well Avidin Plate L15AA-1 L11AA-1

384 Standard 384-well Avidin Plate L25AA-1 L21AA-1
96 High Bind 96-well High-Bind Avidin Plate L15AB-1 L11AB-1
384 High Bind 384-well High-Bind Avidin Plate L25AB-1 L21AB-1

Streptavidin 96 Standard 96-well Streptavidin Plate L15SA-1 L11SA-1

384 Standard 384-well Streptavidin Plate L25SA-1 L21SA-1
96 High Bind 96-well High-Bind Streptavidin Plate L15SB-1 L11SB-1
384 High Bind 384-well High-Bind Streptavidin Plate L25SB-1 L21SB-1

Glutathione 96 High Bind 96-well High-Bind Glutathione Plate L15GB-1 L11GB-1

384 High Bind 384-well High-Bind Glutathione Plate L25GB-1 L21GB-1

Anti-Rabbit 96 SS Standard 96-well Small Spot GAR Plate L45RA-1 L41RA-1

(Goat) 384 Standard 384-well GAR Plate L25RA-1 L21RA-1

Anti-Mouse 96 SS Standard 96-well Small Spot GAM Plate L45MA-1 L41MA-1

(Goat) 384 Standard 384-well GAM Plate L25MA-1 L21MA-1

Protein A 96 High Bind 96-well Protein A Plate L15DB-1 L11DB-1

384 High Bind 384-well Protein A Plate L25DB-1 L21DB-1

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Catalog Numbers

Read Buffers Diluents and Buffers

Description Surfactant Quantity Catalog Number Description Quantity Catalog #
Read Buffer T (4X) + 50 mL R92TC-3 Blocker A Kit 250 mL R93AA-2
Read Buffer T (4X) + 200 mL R92TC-2 1L R93AA-1
Read Buffer T (4X) + 1L R92TC-1

Read Buffer T (4X) - 50 mL R92TD-3 Antibody Diluent 50 mL R50AA-4

Read Buffer T (4X) - 200 mL R92TD-2 200 mL R50AA-2
Read Buffer T (4X) - 1L R92TD-1 1L R50AA-3

Read Buffer P (4X) + 50 mL R92PC-3 Tris Lysis Buffer 50 mL R60TX-3

Read Buffer P (4X) + 200 mL R92PC-2
Read Buffer P (4X) + 1L R92PC-1 200 mL R60TX-2

Read Buffer S (4X) + 50 mL R92SC-3 Tris Wash Buffer (10X) 200 mL R61TX-2
Read Buffer S (4X) + 200 mL R92SC-2 1L R61TX-1
Read Buffer S (4X) + 1L R92SC-1

Read Buffer S (4X) - 50 mL R92SD-3

Read Buffer S (4X) - 200 mL R92SD-2
Read Buffer S (4X) - 1L R92SD-1

Read Buffer G (4X) + 50 mL R92GC-3

Read Buffer G (4X) + 200 mL R92GC-2
Read Buffer G (4X) + 1L R92GC-1

Read Buffer G (4X) - 50 mL R92GD-3

Read Buffer G (4X) - 200 mL R92GD-2
Read Buffer G (4X) - 1L R92GD-1

Ruthenium Products
Conjugated Reporters Labeling Reagents (NHS Ester)
MSD Label Reporter Quantity Catalog Number Description Quantity Catalog Number
SULFO-TAG Anti-Rabbit Antibody (Goat) 50 g R32AB-5 SULFO-TAG 150 nMoles R91AN-1
SULFO-TAG Anti-Rabbit Antibody (Goat) 200 g R32AB-1 SULFO-TAG 500 nMoles R91AN-2
TAG 150 nMoles R91BN-1
SULFO-TAG Anti-Mouse Antibody (Goat) 50 g R32AC-5
SULFO-TAG Anti-Mouse Antibody (Goat) 200 g R32AC-1 TAG 500 nMoles R91BN-2

SULFO-TAG Anti-Human Antibody (Goat) 50 g R32AJ-5

SULFO-TAG Anti-Human Antibody (Goat) 200 g R32AJ-1

SULFO-TAG Streptavidin 50 g R32AD-5

SULFO-TAG Streptavidin 200 g R32AD-1
TAG Anti-Rabbit Antibody (Goat) 200 g R31AB-1

TAG Anti-Mouse Antibody (Goat) 200 g R31AC-1

TAG Streptavidin 500 g R31AD-2

Visit www.mesoscale.com for a complete product listing.

For research use only; not for use in diagnostic or therapeutic procedures. 14


9238 Gaither Road Gaithersburg, MD USA 20877 Phone: 240.631.2522 x4685 Fax: 301.990.2776
Meso Scale Discovery, Meso Scale Diagnostics, MSD, MSD (design), MULTI-ARRAY, MULTI-SPOT, SULFO-TAG and SECTOR are trademarks of Meso Scale Diagnostics, LLC.
2007 Meso Scale Discovery, a division of Meso Scale Diagnostics, LLC. All rights reserved.
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