Date of Completion

2024

Document Type

Honors College Thesis

Department

Molecular Genetics

Thesis Type

Honors College

First Advisor

Dev Majumdar, PhD

Second Advisor

Bruno Martorelli Di Genova, PhD

Third Advisor

Amy Schumer, MD

Keywords

epitope masking, maternal-fetal antibody interaction, SARS-CoV-2, neonatal antibody repertoire, VirScan, PhIP-Seq

Abstract

The ability to determine if there is epitope masking of neonatal antibodies by maternal antibodies would influence knowledge in fetal immunity and disease susceptibility. Here, VirScan, a new application of PhIP-Seq, is applied to mouse dam and pup serum to examine if epitope masking occurs with maternal SARS-CoV-2 antibodies and neonatal SARS-CoV-2 antibodies. VirScan utilizes a peptide library containing the peptides for all of the proteins in the human virome. This VirScan application of PhIP-Seq detects epitopes of human antiviral antibodies. Here the peptide library was expanded to high library diversity. Number of PCR cycles was tested to limit noise from over amplification. It was demonstrated that selection for phage-antibody complexes is not stringent enough, and additional wash steps to remove unbound phage and only pull-down serum-bound phage is beneficial. The VirScan application of PhIP-Seq is a powerful tool for immunological questions, and implementation of methodology adaptations presented here could achieve a successful PhIP-Seq adaptation for numerous uses.

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.

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