Implementing VirScan: a Phage Display Library for Viral Antibody Detection and Characterization
Conference Year
2024
Abstract
Using VirScan, a PhIP-seq peptide library, serological studies looking at antibody repertoires against the entire human virome are possible. By implementing and modifying existing PhIP-seq methods to produce data that is replicable and reliable, such libraries like VirScan can be used to test a multitude of future questions including vaccine efficacy and epitope spread. Through this work, the effectiveness of the existing PhIP-seq method accompanying the VirScan peptide library was tested. The peptide library was amplified and quantified to ensure sufficient diversity was maintained, phage-antibody complexes were formed and precipitated, and captured phage DNA was sequenced. The resulting sequencing data was then analyzed, and peptide hits were calculated using relative z-scores. Through this method, a diverse peptide library was maintained, and peptides hits for Herpesvirus were identified, functioning as a positive control. However, the results were not consistent in sample replicates and positive control samples using monoclonal antibodies did not produce the expected results. This shows a problem of data noise in the existing method and presents the need for protocol adaptation before useful implementation can be achieved.
Primary Faculty Mentor Name
Dev Majumdar
Status
Undergraduate
Student College
College of Agriculture and Life Sciences
Second Student College
Patrick Leahy Honors College
Program/Major
Molecular Genetics
Primary Research Category
Life Sciences
Implementing VirScan: a Phage Display Library for Viral Antibody Detection and Characterization
Using VirScan, a PhIP-seq peptide library, serological studies looking at antibody repertoires against the entire human virome are possible. By implementing and modifying existing PhIP-seq methods to produce data that is replicable and reliable, such libraries like VirScan can be used to test a multitude of future questions including vaccine efficacy and epitope spread. Through this work, the effectiveness of the existing PhIP-seq method accompanying the VirScan peptide library was tested. The peptide library was amplified and quantified to ensure sufficient diversity was maintained, phage-antibody complexes were formed and precipitated, and captured phage DNA was sequenced. The resulting sequencing data was then analyzed, and peptide hits were calculated using relative z-scores. Through this method, a diverse peptide library was maintained, and peptides hits for Herpesvirus were identified, functioning as a positive control. However, the results were not consistent in sample replicates and positive control samples using monoclonal antibodies did not produce the expected results. This shows a problem of data noise in the existing method and presents the need for protocol adaptation before useful implementation can be achieved.