Meso Scale Discovery. Bioprocess Applications

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1 Meso Scale Discovery Bioprocess Applications

2 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 antidrug antibodies in immunogenicity testing. Ultrasensitive cytokine and phosphoprotein kits can enable functional cell based measurements and general potency assays. MSD s instruments are wellsuited 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 CellBased Assays Cytokine & Phosphoprotein Potency Assays CellBased Assays for Antibody Screening Immunogenicity Assays Meso Scale Discovery A division of Meso Scale Diagnostics, LLC Gaither Road Gaithersburg, MD USA Phone: (x4685 for customer service)

3 MSD Technology MSD s electrochemiluminescence detection technology uses TM labels that emit light upon electrochemical stimulation initiated at the electrode surfaces of MULTIARRAY and MULTISPOT microplates. A 1 2 MULTIARRAY Plate B Electrochemiluminescence Features: Minimal background signals and high signal to background ratios the stimulation mechanism (electricity) is decoupled from the signal (light) Proximity only labels bound near the electrode surface are detected, enabling nonwashed assays LIGHT Measured signal is light *Ru(bpy) 2 3 TPA. H Ru(bpy) 2 3 Ru(bpy) 3 3 TPA TPA. e e Luminescence Emitting Light Chemi Chemical Energy Electro Electrochemically Initiated Flexibility labels are stable, nonradioactive, and are conveniently conjugated to biological molecules Emission at ~620 nm eliminating problems with color quenching Counter Electrode Working Electrode Dielectric amplification multiple excitation cycles of each label enhance light levels and improve sensitivity MULTIARRAY and MULTISPOT 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 MULTISPOT Assay Simultaneous measurement of analytes in one well. Capture antibodies are arrayed on the patterned working electrode. Analytes are detected with antibodies labeled with reagent. Dielectric Counter electrode Analyte 1 Analyte 2 Analyte 3 Analyte 4 2

4 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 readymade kits with commercial antibodies or custom kits that incorporate your processspecific 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, antispecies 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 precoated with capture antibody, CHO standards, diluents, and prelabeled detection antibodies. The MSD assay is 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 ELISACHO HCP Assay MSD (200 pg/ml DL)* ELISA (17,000 pg/ml DL) Absorbance (492 nm405 nm) Electrode Surface labeled anticho HCP antibody Host cell protein AntiCHO HCP capture antibody 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 (well plates) Analyte Description SI2400 SI6000 CHO Host Cell Protein Kit CHO Host Cell Protein Kit (1 Plate) K150HNF1 K110HNF1 CHO Host Cell Protein Kit (5 plates) K150HNF2 K110HNF2 CHO Host Cell Protein Kit (20 plates) K150HNF3 K110HNF3 CHO Host Cell Protein Base Kit (5 plates) K150HNA3 K110HNA3 3

5 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 processspecific antibodies. Antibodies can be coated directly on MSD plates or immobilized through common biological linkers (e.g. streptavidinbiotin, antispecies antibodies). The HCP assay shown here was developed using customer antibodies in a bridging format. The assay can also be run in a nowash format with only a modest (2.5 fold) loss in sensitivity. HCP Contamination Assay,000 Washed NoWash labeled antibody Host cell protein Biotinlabeled antibody Biotin Streptavidin Electrode Surface Concentration (ng/ml) MSD (wash) MSD (nowash) LOD (ng/ml) Top (ng/ml) 7,500 7,500 NoWash 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 requirements of the applications, providing a margin of safety. Simple assay formats and wide dynamic ranges reduce time, labor and error by eliminating 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 antihcp antibody is used as both capture antibody (in biotinylated form) and detection antibody ( labeled). The assay is 20 times more sensitive than the comparable ELISA. It also has a larger dynamic range NoWash HCP Contamination Assay,000 MSD (nowash) ELISA LOD (ng/ml) Top (ng/ml) 7,500 5, Concentration (ng/ml) Example Protocol 1. Add 25 µl/well of biotinylated antihost cell protein (HCP) antibody to MSD standard streptavidin plate. 2. Add 25 µl/well of labeled antihcp 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. 4

6 Kits and Assays for Impurities and Residual Contamination Methotrexate (MTX) Contamination Kit Methotrexate is used to select highproducing 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 onewash or nowashed format. This assay incorporates a FITClabeled MTX as the tracer and labeled antifitc as the detection antibody. This format is applicable to other competitive immunoassays. MTX Competitive Format labeled antilabel antibody Free MTX Labeled MTX Directly immobilized antimtx antibody Electrode surface Concentration [ng/ml] Protein A Contamination Kit MSD Protocol 1. Add 20 µl of sample. Add 20 µl of FITClabeled MTX. Incubate 2 hrs. 2. Add 25 µl of labeled antifitc. Incubate 2 hrs. 3. Wash plate 3x with PBST. 4. Add 150 µl of Read Buffer. Read. 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.,000 MSD Protein A Kit γ Globulin Added No lgg labeled antiprotein A antibody Sandwich Format Protein A AntiProtein A antibody Concentration [ng/ml] Electrode surface MSD Protocol 1. Add 20 µl of sample. Incubate 2 hrs. 2. Add 25 µl of labeled antiprotein A. Incubate 2hrs. 3. Wash plate 3x with PBST. 4. Add 150 µl of Read Buffer. Read. 5

7 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. CHOHCP Insulin MTX 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. Protocol antifitc 1. Add 25 µl/well of standards (insulin and CHOHCP) 2 ng/ml of MTXFITC tracer. Incubate for 2 hours. 2. Add 25 µl/well of detection antibodies. Incubate for 2 hours. 3. Wash 3 times with PBST. FITCMTX 4. Add 150 µl/well of 1X Read Buffer T and analyze plate on SECTOR instrument. AntiMTX CHO Contamination Methotrexate Insulin Contamination ECL Counts ECL Counts ECL Counts 1 Concentration [pg/ml] 10 Concentration [pg/ml] 1 Concentration [pg/ml] Detection Limit 300 pg/ml Detection Limit 20 pg/ml Detection Limit 40 pg/ml 6

8 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) For increased sensitivity, MSD offers a sandwich immunoassay that uses a plate coated with Protein A (as the capture species) and labeled F(ab) goatantihuman 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. labeled F(ab) lgg LIGHT LIGHT Protein A Sandwich Assay Concentration [ng/ml] Concentration (ng/ml) ,500 40,000 Protein A Sandwich Assay Average ,165 2,581 8,906 46, , , ,810 StdDev ,634 4,494 13,416 6,817 %CV Protein A plate MSD Protocol 1. Add 25 µl of human IgG standard or sample. Incubate for 2 hours. 2. Wash 3x with PBST. 3. Add 25 µl of F(ab) antihuman detection antibody. Incubate for 1 hour. 4. Wash 3x with PBST. Range of Assay 5 ng/ml 10 µg/ml 5. Add 150 µl of Read Buffer T. Read. 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 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). labeled human IgG Product Protein A,000 Protein A Competition Assay Low Protein A High Protein A Concentration [µg/ml] Concentration (µg/ml) Detection Limit Protein A Competition Assay Average %CV 15, , , , , , µg/ml Protein A Coated 2.1 ng Tracer Concentration 0.31 µg/ml Competition Protocol 1. Add 25 µl of sample. Add 25 µl of labeled human IgG. Incubate 2 hours. 2. Add µl of Read Buffer T (1.5x). Read. 7

9 Assays for Antibody Screening and Production Light Chain and Heavy Chain Immunoassays LIGHT MSD offers assays to detect intact (whole) antibodies. One format uses antiλ or antiκ 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 antiλ capture antibody and a labeled antihuman heavy chain antibody. Purified human immunoglobulins were run along with a sample containing a cocktail of human immunoglobulins. AntiFc Product Antiλ Heavy Chain Light Chain Assay (λ Capture),000 0 lgg lgg1 lgg2 lgg3 lgg Concentration [µg/ml] higg µg/ml higg (mix) , ,986 63, , , ,323 higg ,389 9, ,471 Analyte higg ,003 2,453 11,041 70,397 higg ,150 3, , , , , ,279 higg , , , , , ,288 Multiplex Isotyping Assay 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 isotypespecific capture antibodies with a detection antibody. Purified isotypes were run on the panel in single concentrations to measure the crossreactivity of the assays. The crossreactivities were generally below 2% with the exception of IgG4 and IgM, which crossreacted 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 lgg1 lgg2 lgg3 lgg4 lga lgm lg Concentration [pg/ml] lgg2 Spot lgg1 lgg2 lgg3 lgg4 lga lgm lg Concentration [pg/ml] lgg3 Spot lgg1 lgg2 lgg3 lgg4 lga lgm lg Concentration [pg/ml] Crossreactivity Analyte Spot lgg1 lgg1 % lgg2 2.4% lgg3 0.6% lgg4 0.3% lga 0.0% lgm 0.0% lgg2 lgg3 lgg4 0.3% % 0.5% 0.6% 1.1% 0.0% 0.4% 0.8% % 1.2% 0.1% 0.0% 2.8% 2.7% 1.4% % 0.4% 0.0% lga 0.1% 1.9% 0.5% 0.2% % 0.4% lgm 2.7% 2.7% 1.4% 0.9% 1.2% % lgg1 lgg2 lgg3 lgg4 lga lgm lgg4 Spot lgg1 lgg2 lgg3 lgg4 lga lgm lga Spot lgg1 lgg2 lgg3 lgg4 lga lgm lgm Spot lgg1 lgg2 lgg3 lgg4 lga lgm BSA lg Concentration [pg/ml] lg Concentration [pg/ml] lg Concentration [pg/ml] 8

10 CellBased 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, THP1 and WIL. Assays can be run in well or well plates and can be run with 50015,000 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 nowash format. labeled antilgg Antibody Cell surface protein Cells Carbon electrode MSD Assays to Replace FACS Methods MSD assays can replace FACS analysis for screening antibodies against cellsurface 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. (transfected) (wild type) 7,000 6,000 5,000 4,000 3,000 2, AB1 AB2 AB Concentration of Antibody (µg/ml) Positive 1 Positive 2 Negative 1.2 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. 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 labeled goatantimouse. Incubate for 2 hours. 3. Wash plate 3x with PBS. 4. Add 150 µl of Read Buffer T (without surfactant). Read. 1 Lu, Y., Wong, W.L., Meng, Y.G. (2006) A high throughput electrochemiluminescent cellbinding assay for therapeutic anticd20 antibody selection. Journal of Immunology Methods. 9

11 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 for more information on immunogenicity assay development and a complete listing of materials and reagents. Bridging Immunogenicity Assay: ELISA Comparison,000 MSD ELISA ADA Concentration (ng/ml) labeled drug Antidrug antibody Biotinlabeled drug MSD SA plate MSD assay shows comparable sensitivity to ELISA, with a larger dynamic range and a simple homogenous incubation. Better Free Drug Tolerance Detection of Low Affinity Antibodies Fewer Washes HighThroughput Direct Conjugation of Stable Label Improved Sensitivity Increased Dynamic Range Reduced Sample Volume Higher Binding Capacity MSD Bridging Assay Protocol ELISA Poor No 34 Good Yes s ng/ml 12 logs 25 µl MSD Excellent Yes 1 High Yes 10s ng/ml 34 logs 525 µl 10X More 1. Combine biotindrug, stagdrug and sample in polypropylene plate and incubate 1 hour to overnight. 2. Transfer solution to preblocked standard streptavidin MSD plate. Incubate for 1 hour. 3. Wash assay plate; add Read Buffer T; read plate on SECTOR instrument. Direct Immunogenicity Assays for Protein Drugs ELISA MSD /Background 10 1 MSD ELISA ADA Concentration [ng/ml] antihuman antibody Antidrug antibody Protein drug Neat human serum was used as the sample matrix. The top of the curve is about 1 µg/ml for both formats, but the MSD format is 40 times more sensitive. 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: Better Free Drug Tolerance Detection of Low Affinity Antibodies Fewer Washes HighThroughput Direct Conjugation of Stable Label Improved Sensitivity Increased Dynamic Range Reduction in Reagent Consumption Higher Binding Capacity 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 1 for 1 hour. 3. Wash plate. Add 25 ml of sample. Poor No 35 Good No s ng/ml 12 logs 4. (Optional wash). Add 25 ml of detection antibody. Good Maybe 23 Good No 10s ng/ml 34.5 logs 210 fold 10X More 5. Wash assay plate; add Read Buffer T; read plate on SECTOR instrument. 10

12 Functional Cell Based Assays and Assays for Potency Cytokines MSD cytokine assays are available in single and multianalyte kits. MSD s cytokine protocols are designed to optimize workflow and easeofuse 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. IL4 IFNγ IL5 IL1β IL2 MSD Human TH1/TH2 10 counts IL12p70 IL13 IL8 TNFα IL10 Detection Parameters (pg/ml) Electrode LabeledAb IL8 Capture Ab Colormap display demonstrates different patterns of cytokine responses in different wells of a well 10spot assay plate.,000,000 UltraSensitive Kit IFNγ IL1β IL2 IL4 IL5 IL8 IL10 IL12p70 IL13 TNFα LLOD LLOQ IFNγ IL1β IL IL IL IL IL IL12p IL TNFα Concentration (pg/ml) 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. Mean 30,000 25,000 20,000 15,000 Positive Negative VEGFR2 (ptyr1054/1059) Traditional Western Pos Neg pvegfr2 VEGFR2 MSD Experimental Pos Neg 5, μg lysate per lane 1.3 μg lysate per well Lysates [μg] bfgf 10 5 VEGF antibody Analyte Capture antibody Concentration [pg/ml] Concentration [pg/ml] 11

13 Catalog Numbers Bioprocess Assays Analyte Description SI2400 SI6000 Protein A Protein A Contamination Assay Kit (1 Plate) K150HMF1 K110HMF1 Protein A Contamination Assay Kit (5 plates) K150HMF2 K110HMF2 Protein A Contamination Assay Kit (20 plates) K150HMF3 K110HMF3 Protein A Contamination Assay Base Kit (5 plates) K150HMA3 K110HMA3 CHO Host Cell Protein CHO Host Cell Protein Kit (1 Plate) K150HNF1 K110HNF1 CHO Host Cell Protein Kit (5 Plates) K150HNF2 K110HNF2 CHO Host Cell Protein Kit (20 Plates) K150HNF3 K110HNF3 CHO Host Cell Protein Base Kit (20 Plates) K150HNA3 K110HNA3 Methotrexate Methotrexate Kit (1 Plate) K150H0F1 K110H0F1 Methotrexate Kit (5 Plates) K150H0F2 K110H0F2 Methotrexate Kit (20 Plates) K150H0F3 K110H0F3 Methotrexate Base Kit (20 Plates) K150H0A3 K110H0A3 Protein A, CHO Host Cell Protein BioProcess Duplex Kit (1 Plate) K15149F1 K11149F1 BioProcess Duplex Kit (5 Plates) K15149F2 K11149F2 BioProcess Duplex Kit (20 Plates) K15149F3 K11149F3 BioProcess Duplex Base Kit (20 Plates) K15149A3 K11149A3 Insulin, Methotrexate, CHO Host Cell Protein BioProcess 3plex Kit (1 Plate) K15150F1 K11150F1 BioProcess 3plex Kit (5 Plates) K15150F2 K11150F2 BioProcess 3plex Kit (20 Plates) K15150F3 K11150F3 BioProcess 3plex Base Kit (20 Plates) K15150A3 K11150A3 Immunoassay Development Kits (well Plates) Description PR PR400 SI2400 SI6000 ELISA Conversion Pack I (Uncoated Plates) K13A011 K17A011 K15A011 K11A011 ELISA Conversion Pack II (Antispecies Plates) K13A021 K17A021 K15A021 K11A021 ELISA Conversion Pack III (Avidin/Streptavidin Plates) K13A031 K17A031 K15A031 K11A031 Immunogenicity Development Pack K13A041 K17A041 K15A041 K11A041 Cell Based Assays Development Pack K13A051 K17A051 K15A051 K11A051 Visit for a complete product listing. Contact Information Purchase orders should be faxed or ed to the following queries: TEL: x4685 FAX: customerservice@mesoscale.com 12

14 Catalog Numbers MULTIARRAY Plates for the SECTOR PR Readers Coating Plate Format Binding Capacity Description PR PR400 Bare SS SS well Plate L13XA17XA1 well Small Spot Plate L43XA47XA1 well HighBind Plate L13XB17XB1 well HighBind Small Spot Plate L43XB47XB1 Avidin well Avidin Plate L13AA17AA1 well HighBind Avidin Plate L13AB17AB1 Streptavidin well Streptavidin Plate L13SA17SA1 well HighBind Streptavidin Plate L13SB17SB1 Glutathione well HighBind Glutathione Plate L13GB17GB1 AntiRabbit well GAR Plate L13RA17RA1 AntiMouse well GAM Plate L13MA17MA1 Protein A well Protein A Plate L13DB17DB1 MULTIARRAY Plates for the SECTOR Imagers Coating Plate Format Binding Capacity Description SI2400 SI6000 Bare SS SS well Plate L15XA11XA1 well Small Spot Plate L45XA41XA1 well Plate L25XA21XA1 well HighBind Plate L15XB11XB1 well HighBind Small Spot Plate L45XB41XB1 well HighBind Plate L25XB21XB1 Avidin well Avidin Plate L15AA11AA1 well Avidin Plate L25AA21AA1 well HighBind Avidin Plate L15AB11AB1 well HighBind Avidin Plate L25AB21AB1 Streptavidin well Streptavidin Plate L15SA11SA1 well Streptavidin Plate L25SA21SA1 well HighBind Streptavidin Plate L15SB11SB1 well HighBind Streptavidin Plate L25SB21SB1 Glutathione well HighBind Glutathione Plate L15GB11GB1 well HighBind Glutathione Plate L25GB21GB1 AntiRabbit (Goat) SS well Small Spot GAR Plate L45RA41RA1 well GAR Plate L25RA21RA1 AntiMouse (Goat) SS well Small Spot GAM Plate L45MA41MA1 well GAM Plate L25MA21MA1 Protein A well Protein A Plate L15DB11DB1 well Protein A Plate L25DB21DB1 13

15 Catalog Numbers Read Buffers Description Surfactant Quantity Catalog Number Read Buffer T (4X) Read Buffer T (4X) Read Buffer T (4X) Read Buffer T (4X) Read Buffer T (4X) Read Buffer T (4X) R92TC3 R92TC2 R92TC1 R92TD3 R92TD2 R92TD1 Diluents and Buffers Description Quantity Catalog # Blocker A Kit Antibody Diluent 2 R93AA2 R93AA1 R50AA4 R50AA2 R50AA3 Read Buffer P (4X) Read Buffer P (4X) Read Buffer P (4X) R92PC3 R92PC2 R92PC1 Tris Lysis Buffer R60TX3 R60TX2 Read Buffer S (4X) Read Buffer S (4X) Read Buffer S (4X) R92SC3 R92SC2 R92SC1 Tris Wash Buffer (10X) R61TX2 R61TX1 Read Buffer S (4X) Read Buffer S (4X) Read Buffer S (4X) R92SD3 R92SD2 R92SD1 Read Buffer G (4X) Read Buffer G (4X) Read Buffer G (4X) R92GC3 R92GC2 R92GC1 Read Buffer G (4X) Read Buffer G (4X) Read Buffer G (4X) R92GD3 R92GD2 R92GD1 Ruthenium Products Conjugated Reporters MSD Label Reporter Quantity Catalog Number AntiRabbit Antibody (Goat) AntiRabbit Antibody (Goat) AntiMouse Antibody (Goat) AntiMouse Antibody (Goat) AntiHuman Antibody (Goat) AntiHuman Antibody (Goat) 50 µg 200 µg 50 µg 200 µg 50 µg 200 µg R32AB5 R32AB1 R32AC5 R32AC1 R32AJ5 R32AJ1 Labeling Reagents (NHS Ester) Description Quantity Catalog Number TAG TAG 150 nmoles 500 nmoles 150 nmoles 500 nmoles R91AN1 R91AN2 R91BN1 R91BN2 Streptavidin Streptavidin 50 µg 200 µg R32AD5 R32AD1 TAG AntiRabbit Antibody (Goat) 200 µg R31AB1 TAG AntiMouse Antibody (Goat) 200 µg R31AC1 TAG Streptavidin 500 µg Visit for a complete product listing. R31AD2 For research use only; not for use in diagnostic or therapeutic procedures. 14

16 9238 Gaither Road Gaithersburg, MD USA Phone: x4685 Fax: Meso Scale Discovery, Meso Scale Diagnostics, MSD, MSD (design), MULTIARRAY, MULTISPOT, and SECTOR are trademarks of Meso Scale Diagnostics, LLC Meso Scale Discovery, a division of Meso Scale Diagnostics, LLC. All rights reserved v Jan