Laura Hutchison

Major: Biology

Faculty Advisor: Tanya Miura

Project Title:

Using Deep Mutational Scanning to Understand Antibody Escape by Respiratory Syncytial Virus

Abstract

This project focuses on the monoclonal antibody D25 and its interaction with the RSV fusion glycoprotein (RSV-F). RSV-F mediates viral entry into host cells, while the monoclonal antibody (mAb) D25 neutralizes the ability of RSV-F to function in entry kinetics. RSV-F readily mutates to evade the neutralizing effects of mAbs, resulting in a phenomenon known as antibody escape.

With the help of Twist Biosciences and molecular modeling, a mutational library has been built to use deep mutational scanning (DMS) to analyze 30 amino acid mutations within the D25 epitope of RSV-F. Using molecular modeling predictions of RSV-F mutations that give rise to antibody escape, our lab has previously identified three escape mutations via Illumina sequencing after several passages of RSV in HEp-2 cells with varying concentrations of D25. These mutations, Q202R, N208Y, and N208K were identified, one of which was predicted by molecular modeling.

The use of DMS in this project will aid our understanding of other potential escape mutations within the D25 epitope, leading to the development of a streamlined methodology that can be used in the context of other antibodies related to RSV or other virus-antibody interactions.

Funding: National Science Foundation EPSCoR Research Infrastructure Improvement Program: Track-2, award number OIA-1736253