Molecular Biomarkers & Diagnosis

ISSN: 2155-9929

Open Access

A case study: A robust and fast fluorescent foci-based microneutralization assay using a virus expressing GFP as a key functional biomarker in support of Medimmune clinical studies for the advancement of anti-viral mAb a

4th International Conference on Biomarkers & Clinical Research

July 15-17, 2013 Courtyard by Marriott Philadelphia Downtown, USA

Rui Lin

Scientific Tracks Abstracts: J Mol Biomark Diagn

Abstract :

N eutralizing antibodies against viruses represent a major mechanism of host protection against viral infections. Most if not all marketed vaccines elicit neutralizing antibodies. Currently, neutralizing anti-viral therapeutic monoclonal antibodies are being developed for the treatment and/or the prevention of viral diseases. Standard neutralization assays to assess the viral neutralization activities of antibodies have historically been functional plaque assays. Plaque assays are time-consuming, labor- intensive and challenging to implement in clinical studies especially those involving a large number of patients. Here we described the development and implementation of fast and robust Fluorescent Foci (FFA)-based microneutralization (MN) assays. These assays, using viruses expressing enhanced green fluorescence protein (EGFP), allow for higher throughput, better precision, and shorter assay turn-around time making them suitable for use in large clinical studies. We will first describe the establishment of a novel FFA-based MN assay to detect and quantify neutralizing antibodies against EBV. This assay uses EBV-GFP and an engineered epithelial cell line. All assay conditions impacting assay performance were optimized and the assay was automated using liquid handling and high-content imaging systems. The robustness and precision of the optimized assay were demonstrated using serum samples from mice, rabbits and humans (n > 600). In addition, a companion EBV-specific IgG ELISA assay was developed. Significant correlation (r2 = 0.89) between both assays was demonstrated using 358 rabbit serum samples. A MN assay was similarly developed for Respiratory Syncytial virus (RSV) to support the development of MedImmune?s anti-RSV therapeutic mAbs and RSV vaccine candidates. The 2-year assay control trending of RSV MN assay will be presented to demonstrate the robustness and precision of this assay

Biography :

Rui Lin received his Ph.D. from Cornell University. He completed his postdoc work in UC San Francisco and Genentech. His past work included signal transduction and anti-angiogenesis research, drug discovery and biomarker studies. He is currently a Scientist in Translational Medicine at MedImmune working on vaccine and monoclonal therapeutic antibodies.

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