(467d) Advancing Next-Generation Sequencing Based Methods for Antibody Discovery and Engineering

A major part of the advancement of the field of systems immunology has been the emergence of next-generation sequencing (NGS); specifically the ability to use NGS for large-scale analysis of antibody variable gene repertoires. Here, I will provide an overview of this exciting new research field, which has applications related to answering basic questions of antibody diversity, development, and evolution; as well as applications for immune-diagnostics and immunotherapeutics.  I will primarily focus on our research group's interests in applying NGS of antibody repertoires for the discovery and engineering of monoclonal antibody (mAb) therapeutics. We have developed a number of experimental and computational methods to help advance NGS of antibody repertoires. These include a high-throughput bioinformatic pipeline for NGS analysis and annotation. Also, we have evaluated the robustness and reproducibility of antibody NGS sequencing methods using triplicate sample sequencing and mathematical ecology-based modeling. Finally, we have assessed experimental methods used to construct NGS antibody libraries, focusing on primer bias and the impact of adaptor addition by PCR or ligation. Our work on methods development should help researchers pursue their own goals related to antibody NGS and help advance monoclonal antibody discovery and engineering.