Applications of the Corning® Epic® System for Label-Free Antibody and Antigen Detection

Arron Xu, Anthony G. FrutosCorning Incorporated, Life Sciences, One Science Center Drive, Corning, NY 14831

The Corning® Epic® System is a high-throughput, label-free detection platform that consists of SBS-standard 384-well microplates with optical sensors inside each well, an HTS-compatible microplate reader and a set of label-independent assay protocols. The Epic® System is applicable to both biochemical and cell-based assays, and enables high-throughput screening of “intractable” targets.

Corning® Epic® System Concept

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Microplate Microplate Reader Assay Dataª 384-well formatª Optical biosensor in each well

ª ≥ 40,000 wells/8 hrsª Compatible with HTS automation

ª Manipulated and analyzed by customer

ª Sensitivity of 5pg/mm2

(300Da drug to 75kDa target)

The Corning® Epic® System technology is a novel high-throughputlabel-free de-tection technology developed for both biochemical and cell-based assays in drug discovery. The study presented here highlights the applications of this technol-ogy for the detection of antibody-antigen and antibody-antibody binding assays in complex biological samples. Two model detection systems are discussed here: 1) streptavidin and antibody interaction; 2) F(ab’)2 and antibody interaction. Re-sults demonstrate the feasibility of detecting streptavidin by an anti-streptavidin antibody immobilized on the Epic® 384-well biosensor. Such a detection system can also be ‘flip-flopped’ by immobilizing streptavidin on the Epic® biosensor, thus providing an alternative assay mechanism that otherwise is limited by conventional label-dependent technologies. We also report the detection of a target antibody by sensor-immobilized F(ab’)2. Detection of protein binding targets or partners in bovine serum was successful. These results demonstrate the Epic® system as a high-throughput label-free platform suitable for affinity based detection of pro-tein-protein interaction.

AbstractCorning® Epic® System technology is a novel high-throughput label-free detection technology suitable for both biochemical and cell-based assays for drug discov-ery. Potential leads are discovered by detection of the binding of small drug com-pounds to biological targets that leads to change of the refractive index. This in turn changes the measured wavelength of the refractive light by the Epic® resonant waveguide grating biosensor.

Conventional technologies for high-throughput protein/protein binding assay center around label-dependent technologies (e.g. ELISA) technologies. These technologies are limited by the requirement of secondary labeling. We focus in the preliminary study on exploring the feasibility of using the Epic® System for detec-tion of antibody-antigen and antibody/antibody binding.

Introduction

Part A: Immobilized F(ab’)2 Goat anti-Mouse IgG (Fc g) Capture of Mouse IgG1, k

Part B: Antibody and Streptavidin Interaction

Principle of Detection Immobilized F(ab’)2 Capturing of Mouse IgG1, k in PBS

Immobilized F(ab’)2 Detection of Mouse IgG1, k in 5% Fetal Bovine Serum

Summary of Immobilized F(ab’)2 Detection of Mouse IgG1, k

Immobilized F(ab’)2 Detection of Mouse IgG1 k in PBS

Immobilized F(ab’)2 Detection of Mouse IgG1, k in Bovine Serum up to 90% (v/v)

Principle of Detection Principle of Detection

Immobilized Streptavidin Detection Anti-streptavidin Antibody in up to 85%(v/v) Serum

Anti-streptavidin Antibody Detection in Serum by Immobilized Streptavidin

Summary of Immobilized Streptavidin Detection of Antibody

For Correspondence: epic@corning.comPresented at LabAutomation, January 2007