Development and Application of Statistical Analysis for Epitope Mapping

• Activity Number: 172
• Type: Contributed
• Date/Time: Monday, August 2, 2010 : 10:30 AM to 12:20 PM
• Sponsor: Biopharmaceutical Section
• Author(s): Cheng Su*+ and Ning Weng
• Companies: Amgen Inc. and Amgen Inc.
• Keywords: epitope mapping ; automated analysis ; non linear curve fitting ; application

Abstract

We aimed at developing standardized statistical analysis to aid epitope mapping - the process of locating regions in antigen/protein that interact with antibodies. The experimental method described herein is arginine scanning mutagenesis in which up to 100 mutant proteins, each with a single-site arginine mutation, were made and their bindings to antibodies tested in bead-based multiplex assay. The mutant binding curves were compared to that of the wild type; mutations that caused significant changes in binding are considered as part of the interaction epitope. In collaboration with researchers, we developed an automated analysis workflow that includes estimation of bead region variability, data QC, nonlinear curve fitting and QC, and curve comparisons. The application of this workflow has led to significant time saving and better results.