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

Cell Rep Methods. 2024 Dec 16;4(12):100934. doi: 10.1016/j.crmeth.2024.100934.

Abstract

In this work, we developed PolyMap (polyclonal mapping), a high-throughput method for mapping protein-protein interactions. We demonstrated the mapping of thousands of antigen-antibody interactions between diverse antibody libraries isolated from convalescent and vaccinated COVID-19 donors and a set of clinically relevant SARS-CoV-2 spike variants. We identified over 150 antibodies with a variety of distinctive binding patterns toward the antigen variants and found a broader binding profile, including targeting of the Omicron variant, in the antibody repertoires of more recent donors. We then used these data to select mixtures of a small number of clones with complementary reactivity that together provide strong potency and broad neutralization. PolyMap is a generalizable platform that can be used for one-pot epitope mapping, immune repertoire profiling, and therapeutic design and, in the future, could be expanded to other families of interacting proteins.

Keywords: CP: Biotechnology; CP: Immunology; PolyMap; SARS-CoV-2 surface antigen; antibody-antigen interaction; cell surface protein; high-throughput screening; polyclonal antibody profiling; protein-protein interaction; ribosome display; single-cell analysis; viral neutralization.

MeSH terms

  • Antibodies, Neutralizing / immunology
  • Antibodies, Viral* / immunology
  • Antibody Specificity
  • COVID-19* / immunology
  • Epitope Mapping / methods
  • High-Throughput Screening Assays / methods
  • Humans
  • Microfluidics / methods
  • Peptide Library
  • Ribosomes / metabolism
  • SARS-CoV-2* / immunology
  • Spike Glycoprotein, Coronavirus* / chemistry
  • Spike Glycoprotein, Coronavirus* / genetics
  • Spike Glycoprotein, Coronavirus* / immunology
  • Spike Glycoprotein, Coronavirus* / metabolism

Substances

  • Spike Glycoprotein, Coronavirus
  • Antibodies, Viral
  • spike protein, SARS-CoV-2
  • Peptide Library
  • Antibodies, Neutralizing

Supplementary concepts

  • SARS-CoV-2 variants