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MSD Confidential and Proprietary5/26/2016
Slide 1
MESO SCALE DISCOVERY PLATE BASEDELECTROCHEMILUMINESCENT TECHNOLOGY
Jimmy D. Page, Ph.D.
Scientific Services
MSD Confidential and Proprietary5/26/2016
Slide 2
MSD® Business Overview
In business since 1995 (20 years)• Pharma, Biotech and Life Sciences
• Government
• Clinical
Over 1500 Units Worldwide• All major pharma and biotechs
• Clinical Research Organizations, andAcademic Core Labs.
• Personalized Medicine Companies
• Government labs
• Most customers with multiple instrumentsper site.
Long-term corporate commitment• ISO 9001:2008 certified QMS
• ISO 13485:2003 certified QMS for designand development.
• Expansion into a validated manufacturingfacility mid-2014
Research to late stage applications• Clinical biomarkers
• Personalized Medicine / CompanionDiagnostic
• Immunogenicity
• Serological markers / Vaccine
• Bioprocess and QC
MSD Confidential and Proprietary5/26/2016
Slide 3
Electrochemiluminescence
Electro-
Chemi-
Luminescenc-SULFO-TAGTM Label
Ruthenium (II) tris-bipyridine NHS ester
Robust Detection
Small molecule <1000 Da
Stable, Non-radioactive
No auto-electrochemiluminescence
Emission at 620 nm
Elimination of color quenching.
Low backgrounds: stimulationmethod (electricity) is decoupledfrom the signal (light).
Sensitive: multiple excitationcycles can amplify signals toenhance light levels.
Large dynamic range: generationof light not absorbance.
MSD Confidential and Proprietary5/26/2016
Slide 4
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 electrode surfaceare detected.
Flexible: Non-washed assays
Electrode surface can be coated directly
MULTI-ARRAY® Plate
SULFO-TAGTM
labels
MSD Confidential and Proprietary5/26/2016
Slide 5
MSD Multiplex Plates – Catalog or Custom
Workingelectrode
“spot”
Counterelectrode
Dielectric
Support
1 2
A
B
Workingelectrode
“spot”
Counterelectrode
Dielectric
1 2
A
B
384 well plate:1 or 4 spots/well
4
7
10
96 well plate1, 4, 7 or 10 spots/well
1
Manufactured using deposition of conductive and insulating layers
similar to computer circuit board manufacturing.
Multiple electrical circuits within the same well.
Each circuit is spatially independent.
Biospecific reagents are deposited on each circuit or “spot.”
Multiplexed assays are possible through the specificity of the capture
reagents and the ability to resolve images.
MSD Confidential and Proprietary5/26/2016
Slide 6
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; Barcode for automatedread
No Operator Calibration
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 (21 CFR Part 11)
QC features in 4.0 Software.
MSD Confidential and Proprietary5/26/2016
Slide 7
The MSD Advantage; Dynamic Range
Human TNFa
0.1 1 10 100 1000 10000
Normal Human
Disease Human
Typical ELISA
MSD Assay
Sensitive ELISA
Concentration (pg/ml)0.1 100
MSD V-PLEX TNFa Assay
High Sensitivity
Large Dynamic Range
High Precision
Low Background
Simple Protocols
Reduced Matrix Effects
0.01 0.1 1 10 100 100010
100
1000
10000
100000
1000000
TNF- Concentration (pg/mL)
Sig
nal
MSD Confidential and Proprietary5/26/2016
Slide 8
The MSD Advantage: Multiplexing
• Large dynamic range assays (3-4 logs)
• Detection limits between 0.1 and 10pg/ml.
• Low sample volume (1-25 µl)• Can use cell supernatants, serum,
plasma, or whole blood.
0.01 0.1 1 10 100 1000 10000100
1000
10000
100000
1000000
IFN
IL-1
IL-2
IL-4
IL-6
IL-8
IL-10
IL-12p70
IL-13
TNF
Concentration (pg/mL)
Sig
nal
Proinflammatory Panel 1(human) V-PLEX Kit
MSD Confidential and Proprietary5/26/2016
Slide 9
Compatibility with Complex Samples
Data shown is from one spot of a 7-plex plate,precious sample is conserved and measured atmuch higher throughput.
Ability to analyze complex matrices including:
Serum /Plasma
Cell lysates /Tissue Homogenates
BAL
Nasal lavage
Sputum
Synovial fluid
Breath Condensate
CSF
Urine
Tumor extracts
Blood spots
Vaginal secretions
Tears
Validated by clinical labs.
IL-1b (Human Placenta)
covariance r = 0.9999557
0.1
1
10
100
1000
10000
0.1 1 10 100 1000 10000
MSD
EL
ISA
Brigham and Women’sHospital
Harvard MedicalSchool
Fichorova et. al,“Trichomonas vaginalis Lipophosphoglycan Triggers a Selective Upregulation of Cytokines by Human FemaleReproductive Tract Epithelial Cells” INFECTION AND IMMUNITY, Oct. 2006, p. 5773–5779
MSD Confidential and Proprietary5/26/2016
Slide 10
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
MSD Confidential and Proprietary5/26/2016
Slide 11
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)
MSD Confidential and Proprietary5/26/2016
Slide 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
MSD Confidential and Proprietary5/26/2016
Slide 13
Benefits of MSD Technology
Larger dynamic range – reduces requirement for sample dilutions, fewerretests due to samples being out of range.
High sensitivity
Reduced samples volumes conserve on reagents and ultimately cost. Manyof the serum-based assays are done at a fraction of the sample, thus savingprecious samples.
Complex Matrices are easily run. Higher matrix tolerance improves assaysensitivity
Improved workflow because there is a significant reduction in the number ofassay and wash steps. Running multiple assays in a given experiment canincrease throughput by more than 10 fold.
Flexibility in building assays in multiple different formats and rapid assaydevelopment.
MSD Confidential and Proprietary5/26/2016
Slide 14
Contact Details:Jimmy Page, Ph.D. Melanie MunozScientific Support Account ManagerTel: 301-525-8424 Tel: 813-484-1171E-mail; jpage@mesoscale.com E-mail; mmunoz@mesoscale.com
MSD Confidential and Proprietary5/26/2016
Slide 15
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 500 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
MSD Confidential and Proprietary5/26/2016
Slide 16
10-Spot vs. Single Spot Performance
Same performance between single-spot and 10-spot (>50 assays validated)
Signal differences may occur, but sensitivity and quantitative range is similar
Individual assays may have slightly better sensitivity because of lowerbackground
0.01 0.1 1 10 100 100010
100
1000
10000
100000
1000000
10-plex
Singleplex
TNF- Concentration (pg/mL)
Sig
nal
Limits of Detection of40 V-PLEX Cytokine Assays
0.01 0.1 1 100.01
0.1
1
10
Individual Assay LLOD (pg/ml)10-P
lex
Ass
ay
LLO
D(p
g/m
l)
®
®
10-Spot Plate
Small Spot Plate
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