meso scale discovery plate based ... - emory university

MSD Confidential and Proprietary5/26/2016

Slide 1

MESO SCALE DISCOVERY PLATE BASEDELECTROCHEMILUMINESCENT TECHNOLOGY

Jimmy D. Page, Ph.D.

Scientific Services


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


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.


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


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.


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.


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


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


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


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


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)


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


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.


Slide 14

Contact Details:Jimmy Page, Ph.D. Melanie MunozScientific Support Account ManagerTel: 301-525-8424 Tel: 813-484-1171E-mail; [email protected] E-mail; [email protected]


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


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

meso scale discovery plate based ... - emory university

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