High-throughput specificity profiling of antibody libraries using ribosome display and microfluidics

Area of Science:

• Immunology
• Biotechnology
• Molecular Biology

Background:

• Understanding antigen-antibody interactions is crucial for vaccine development and therapeutic design.
• Characterizing immune responses to viral variants like SARS-CoV-2 requires high-throughput mapping methods.

Purpose of the Study:

• To develop and validate PolyMap, a novel high-throughput platform for mapping protein-protein interactions, specifically antigen-antibody interactions.
• To profile antibody repertoires from COVID-19 donors against SARS-CoV-2 spike variants.
• To identify and select potent, broadly neutralizing antibodies for therapeutic applications.

Main Methods:

• Developed PolyMap (polyclonal mapping), a high-throughput method for mapping protein-protein interactions.
• Applied PolyMap to analyze thousands of antigen-antibody interactions between antibody libraries from COVID-19 donors and SARS-CoV-2 spike variants.
• Utilized binding data to select antibody mixtures with complementary reactivity for enhanced neutralization.

Main Results:

• Successfully mapped thousands of antigen-antibody interactions, identifying over 150 antibodies with distinct binding patterns.
• Observed broader antibody binding profiles, including reactivity against the Omicron variant, in recent donor repertoires.
• Demonstrated the selection of antibody mixtures providing strong potency and broad neutralization.

Conclusions:

• PolyMap is a generalizable platform for epitope mapping, immune repertoire profiling, and therapeutic antibody design.
• The method facilitates the identification of antibodies with broad reactivity, including against emerging viral variants.
• PolyMap can accelerate the development of effective antibody-based therapeutics and vaccines.