msd business overview - emory university10/11/2016 neurodegeneration assays © 2013 meso scale...

10/11/2016

Neurodegeneration Assays

© 2013 Meso Scale Diagnostics, LLC. All rights reserved.

MSD® Business Overview• In business since 1995 (18+ years)

– Pharma, Biotech and Life Sciences

– Biodefense and Clinical

• Worldwide customer base withover 1300 units

– All major pharma and biotechs

– Clinical Research Organizations, andAcademic Core Labs.

– Personalized Medicine Companies

– Government labs

– More than 1300 active units with mostcompanies with multiple instrumentsper site.

• Long-term corporate commitment– ISO 9001:2008 certified

– ISO 13485:2003 certified for designand development.

– Expansion into a validatedmanufacturing facility

• From research to late stageapplications

– Clinical biomarkers

– Personalized Medicine / CompanionDiagnostic

– Immunogenicity

– Serological markers / Vaccine

– Bioprocess and QC

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MSD Platform

Electrochemiluminescence• Large Dynamic Range• High Sensitivity• Compatible with Complex

Sample Matrices• Small Sample Volumes• Robust Plate Based

Immunoassays

Robust Instrumentation• Multiplex Enabled• Non-Fluidics Based

Instrument.• CCD Imaging System• Validated in Regulated

Environment• 96 and 384-well Plates• 1 minute read time per plate

Available Kits• Over 400 Kits (single analyte

and multiplex)• Simple Workflow

Flexible Assay System• Custom/ novel multiplexes• Labeling Reagents• Secondary Reporters• Assay development services

Development Services• Applications Scientist Support• Contract Manufacturing• Assay development services

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Electrochemiluminescence Implementation

• Simple to Use

– 96 and 384-well plate formats withelectrodes at the bottom of the well.

– Method similarity to ELISA. Detectionconjugated.

– Excitation through plate bottom

• Fast: Excitation of less than 2 sec.Measured through a CCD camera.

• Low Background:

– Background images subtracted

– Only labels near the electrodesurface are detected.

• Flexible:

– Non-washed assays

– Electrode surface can be coateddirectly

MULTI-ARRAY® Plate

SULFO-TAGTM

labels

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Multiplex Assays

• Multiple spots within each well (an array)

• Each spot is an individual assay for a specific analyte

• Capture reagents are deposited on each spot

• Add as single sample and labeled detection reagents

• Measure light emitted from each spot (CCD camera)

• Light from each spot is related to the amount of each analyte in sample

Workingelectrode

“spot”

Counterelectrode

Dielectric

Support

1 2

A

B

Workingelectrode

“spot”

Counterelectrode

Dielectric

1 2

A

B

384 well plate:1 or 4 assays/well

4

7

10

96 well plate1, 4, 7 or 10 assays/well

1

Positional Multiplexing, notbased upon differentwavelengths of fluorophores.

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Instrumentation and Software

• High Sensitive Imaging Detection Systems

– Ultra-low noise CCD camera with 6 logs ofdynamic range

– Automated image analysis with backgroundsubtraction.

• Simple and Robust:

– No Fluidics;

– 2 Clicks to Read

– No Operator Calibration

– Barcode for automated read

– 1 to 3 minutes to read a complete plate

– Integrated into automation platforms

• Regulated Environments:

– Electronic plate for verification

– IQ/OQ/PQ Package

– GLP Compliant Software (CFR21 Part 11)

– QC features in 4.0 Software.

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MSD’s Biomarker Mission

RESEARCH

TRADITIONALCLINICAL

DIAGNOSTICS

ELISA 5000+ Analytes

Variable QualityRelative Quantitation

Abbott, Roche, Siemens

100 Analytes

Diverse Species

Rigorous Quality

Absolute Quantitation

Humans

CLINICALBIOMARKERS

5000+ Analytes Diverse Species Absolute QuantitationRigorous Quality

High Throughput

Limited Sample

Single Marker

Multi-Analyte

Regulatory Approval

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What if your assay is not in the catalog?

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ASSAY DEVELOPMENT TOOLS

PLATES• Bare (uncoated)• Streptavidin/Avidin• Anti-species (mouse, rabbit)• Glutathione• Protein A

TECHNICAL ASSISTANCE/ADVICE• Team of applications scientists• R&D Department

REAGENTS

Buffers

Blockers

DETECTION

Sulfo-Tag NHS Ester

Streptavidin Sulfo-Tag

Anti-species Sulfo-TagAntibodies

CUSTOM COATING SERVICES

Your antibodies coated ontoplates using MSDmanufacturing capabilities

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Open platform to easily develop or transfer assays

• Matched antibody pairs are readilyavailable through a variety of sources.

1 100 10000

100

1000

10000

100000

+61x10 MSD

ELISA

Range ofELISA

Capture antibody

Sample

Biotin-anti-Targetdetection antibody

StreptavidinSULFO-TAG™

Capture antibody

Sample

Biotin-anti-Targetdetection antibody

Streptavidin HRP

Notes:

Reagents used in ELISA can be useddirectly but at lower amounts andvolumes.

Antibodies can be easily adsorbed to thecarbon electrode surface at neutral pHwithout the use of coupling chemistries.The carbon provides a high surfacecapacity to capture a lot of material.

Direct labeling of the detection antibodyis simple.

Typical sensitivity (pg/ml to ng/ml)

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ELISA Conversion

• The sCD30 assay used an in-house monoclonal antibody for capture and abiotinylated polyclonal antibody from R&D Systems for the detection antibody.

• The ST2 assay used a matched antibody pair from R&D Systems with abiotinylated detection antibody.

• The sensitivity of the ELISA assays was stated to be ~ 800 pg/mL for thesCD30 assay and ~ 250 pg/mL for the ST2 assay.

• This sensitivity was not believed to be sufficient to measure endogenouslevels.

• Conversion of the sCD30 assay from the ELISA to MSD using the sameantibody pair resulted in an improvement in the assay sensitivity ofapproximately 15-fold (from ~800 pg/mL to < 50 pg/mL).

• Similarly, the conversion of the ST2 ELISA to MSD gave about a 10-foldincrease in sensitivity (~ 250 pg/mL to ~ 20 pg/mL).

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Assay I Dynamic Range vs ELISA

Meso Scale Discovery © 2012 160218-JECSlide 12

U-PLEX® is a New Technology Platform from MSD®

U-PLEX enables you to design and build your ownpersonalized multiplex assays any time you choose

Open platform: compatible with antibodies, proteins,peptides, carbohydrates, nucleic acids, andoligonucleotides

Use your own antibodies or reagents provided by MSD

Easy workflow and simple protocol similar to ELISA

No spotting required

Compatible with all MSD instruments and software


U-PLEX® is a New Technology Platform from MSD®

U-PLEX enables you to design and build your ownpersonalized multiplex assays any time you choose

Open platform: compatible with antibodies, proteins,peptides, carbohydrates, nucleic acids, andoligonucleotides

Use your own antibodies or reagents provided by MSD

Easy workflow and simple protocol similar to ELISA

No spotting required

Compatible with all MSD instruments and software


U-PLEX Assays and Assay Development

Human

• 51 Assays• 12 popular assay combinations*

Non Human Primate

• 36 Assays• 7 popular assay combinations

Mouse

• 14 Assays• 2 popular assay combinations

Single analyte detection with U-PLEX AntibodySets on MSD GOLD™ Small Spot Streptavidin

Open Spots

• Develop in-house assays• Combine with MSD assays for ultimate

flexibility

Assays Assay Development

* Popular combinations of analytes grouped into separate Cat. Nos. for ordering convenience


U-PLEX Fundamentals

• U-PLEX plates

• Up to 10 spots, each spot activated tocapture a specific U-PLEX linker(represented by color)

• U-PLEX linkers

• 10 linkers, each designed to bind to aspecific spot on the U-PLEX plate

• U-PLEX reagents

• Assay reagents (e.g., MSD GOLDSULFO-TAG™ NHS-Ester)

• Antibodies, Calibrators

• Diluents

• Read Buffer T (4X)


Build Personalized Multiplexes in 3 Simple Steps

• Couple:• Combine antibodies and linkers

• No purification of reagents

• No complex washing

• Combine coupled antibodies

• Coat:• Simple 50 mL addition to plates

• Incubate for 1 hour to overnight

• Plates are ready to use or store for 7 days

• Complete (Run Assay):• Add 25–50 mL of samples for 1 hour

• Add 25 mL of detection antibody for 1 hour

• Read


U-PLEX Linker Coupling

Biotin-binding domain

Matched linkerand spot

One spot of a plate


Combine U-PLEX-Coupled Antibodies

• Select your analytes and choose theappropriate antibodies

• Combine the antibodies to make one solution

• Up to 10 U-PLEX-coupled antibodies canbe pooled

• Do not combine U-PLEX-coupledantibody solutions that share the samelinker

• Final solution is used to coat plates

• One step, not 10

• Antibodies bind to their spots on the plate fromsolution


Multiplexing with U-PLEX Products

U-PLEX plates

10-spot plates activated to capturespecific U-PLEX linkers (represented bycolor)

U-PLEX Linker-Coupled Capture

Biotinylated molecules coupled to

U-PLEX linkers

Add 50 mL of solution and the coupledantibodies bind to the correct spot

U-PLEX Plate U-PLEXLinker-CoupledAntibody


U-PLEX has All the Advantages of MSD’s ECL Technology


U-PLEX Conserves Precious Reagents

Antibody used is less than 2 mg per plate

Reagent costs can be reduced by up to 50-fold from typical ELISAs

Technology/Format Antibody Coating Antibody per PlateELISA 100 mL per well at 1–10 mg/mL 10–100 mg

Luminex1–125 mg per 5x106 beads; 5000 beads per

well 0.1–12.5 mg

MSD Streptavidin 25–50 ml per well at 0.1–1 mg/mL 0.25–5 mgMSD Standard Plates

Hand Spotting 5 ml per well at 2–50 mg/mL 1–25 mgSolution 50 ml per well at 2–20 mg/mL 10–100 mg

U-PLEX 200 ml per plate at 10 mg/mL <2 mg


U-PLEX Human Combinations: Grouped for Ordering Convenience

Name Analytes

U-PLEX Biomarker Group 1 (hu) 40-Plex All 40 Assays from Group 1A launch

U-PLEX TH1/TH2 Combo (hu) IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-8, IL-10, IL-12p70, IL-13, TNF-α

U-PLEX Proinflam Combo 1 (hu) IFN-γ, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13, TNF-α

U-PLEX Proinflam Combo 2 (hu) GM-CSF, IFN-γ, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70

U-PLEX Proinflam Combo 3 (hu) IFN-γ, IL-1β, IL-6, TNF-α

U-PLEX Proinflam Combo 4 (hu) IL-1β, IL-6, IL-8, TNF-α

U-PLEX Cytokine Combo 1 (hu) GM-CSF, IL-1α, IL-5, IL-7, IL-12/IL-23p40, IL-15, IL-16, IL-17A, TNF-β, VEGF-A

U-PLEX Chemokine Combo 1 (hu) Eotaxin, Eotaxin-3, IL-8, IP-10, MCP-1, MCP-4, MDC, MIP-1α, MIP-1β, TARC

U-PLEX Th17 Cytokine Combo 1 (hu) IL-17A, IL-17E/IL-25, IL-17F, IL-21, IL-22, IL-23, IL-27, IL-31, IL-33

U-PLEX Th17 Cytokine Combo 2 (hu) IFN-γ, IL-1β, IL-6, IL-10, IL-17A, IL-17E/IL-25, IL-17F, IL-21, IL-22, TNF-α

U-PLEX Biomarker Group 1 (hu) 40-Plex CTACK, ENA-78, Eotaxin, Eotaxin-3, Fractalkine, G-CSF, GM-CSF, IFN-α2a, IFN-γ, IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12/IL-23p40, IL-12p70, IL-13, IL-15, IL-16, IL-17A,IL-18, IP-10, I-TAC, MCP-1, MCP-4, MDC, MIP-1α, MIP-1β, MIP-3α, MIP-3β, SDF-1α, TARC,TNF-α, TNF-β, TPO, VEGF-A

U-PLEX Biomarker Group 1 (hu) 51-Plex, SECTOR All 51 Human Assays


U-PLEX NHP & Mouse Combinations: Grouped for Ordering Convenience

Name Analytes

U-PLEX Proinflam Combo 1 (NHP) IFN-γ, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12/IL-23p40, TNF-α

U-PLEX Cytokine Combo 1 (NHP) GM-CSF, IL-1α, IL-5, IL-7, IL-12/IL-23p40, IL-15, IL-16, IL-17A, TNF-β, VEGF-A

U-PLEX Chemokine Combo 1 (NHP) Eotaxin, Eotaxin-3, IL-8, IP-10, MCP-1, MCP-4, MDC, MIP-1α, MIP-1β, TARC

U-PLEX Th17 Cytokine Combo (NHP) IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-17A, TNF-α

U-PLEX TH1/TH2 Combo (NHP) IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-8 (CXCL8), IL-10, IL-12p70, TNF-α

U-PLEX Biomarker Group 1 (NHP) 30-Plex CTACK, Eotaxin-3, Fractalkine, G-CSF, GM-CSF, IFN-γ, IL-1β, IL-2, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12/IL-23p40, IL-15, IL-16, IL-17A, IL-18, IP-10, I-TAC, MCP-1, MCP-4, MDC, MIP-1α, MIP-1β,MIP-3β, TARC, TNF-β, TPO, VEGF-A

U-PLEX Biomarker Group 1 (NHP) 36-Plex All 36 NHP Assays

Name Analytes

U-PLEX Biomarker Group 1 (ms) 14-Plex All 14 Mouse Assays

U-PLEX TH1/TH2 Combo (ms) IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-8, IL-10, IL-12p70, IL-13, TNF-α

NHP

Mouse


U-PLEX is an Open Platform Supporting Many Applications

• Antibody Development

• Antibody Screening

• Biomarker Feasibility

• Biomarkers in Complex Matrices

• Homogenous Assays

• Multiplex Immunogenicity/PK

• Nucleic Acid Assays

• Serology and Autoantibody

• Vaccine Studies

biotinylated captureantibody

analyte

SULFO-TAG™detectionantibody

detectionantibody

SULFO-TAG anti-speciesantibody

biotinylatedprotein

SULFO-TAG anti-speciesantibody

antibody/drugbiotinylatedpeptide,carbohydrate

antibody/drug

biotinylatedprobe

SULFO-TAG probe

Oligonucleotide target


S-PLEX

S-PLEX takes MSD’s proven electrochemiluminescence technology to thenext level of sensitivity. These new assays allow you to quantitate previouslyunmeasurable baseline samples with fg/mL sensitivity and a wide dynamicrange while preserving your precious samples. S-PLEX assays are compatiblewith MSD’s market leading SECTOR and QuickPlex instruments, offeringthe ability to leverage our widely accepted technology platform, andeliminating the need to purchase new instrumentation or validate newtechnology.

Highlights◾1000x more sensitive than standard ELISAs◾Proven technology platform◾Simple, robust methods and multiplex capability◾Runs on existing SECTOR and QuickPlex instruments◾Wide dynamic range


S-PLEX IL-6 Assay


S-PLEX Assay Menu

Currently available as a service

Cardiac Troponin IHIV p24IL-2IL-4IL-6IL-10Tau

Coming soon as a service

Complexed PSAFree PSAGM-CSFIL-1βIL-5IL-17AIL-21TNFα

In development – please inquire

Aβ42Aβ40Aβ38Cardiac Troponin TG-CSFIFNγIL-22IL-33IL-12p70IL-23IL-31TSLP


Contact Details:

Jimmy Page, Ph.D. Melanie Munoz

Scientific Support Account ManagerTel: 301-525-8424 Tel: 813-484-1171E-mail: [email protected] E-mail; [email protected]

msd business overview - emory university10/11/2016 neurodegeneration assays © 2013 meso scale...

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