Protocol for constructing large size human antibody heavy chain variable domain (VH) library and selection of SARS-CoV-2 neutralizing antibody domains

STAR Protoc. 2021 Sep 17;2(3):100617. doi: 10.1016/j.xpro.2021.100617. Epub 2021 May 29.

Abstract

This protocol is a comprehensive guide to phage display-based selection of virus neutralizing VH antibody domains. It details three optimized parts including (1) construction of a large-sized (theoretically > 1011) naïve human antibody heavy chain domain library, (2) SARS-CoV-2 antigen expression and stable cell line construction, and (3) library panning for selection of SARS-CoV-2-specific antibody domains. Using this protocol, we identified a high-affinity neutralizing human VH antibody domain, VH ab8, which exhibits high prophylactic and therapeutic efficacy. For complete details on the use and execution of this protocol, please refer to Li et al. (2020).

Keywords: Antibody; Cell culture; High Throughput Screening; Immunology; Microbiology; Molecular Biology; Protein expression and purification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antibodies, Neutralizing / immunology*
  • Antibodies, Viral / immunology*
  • Base Sequence
  • COVID-19 / immunology*
  • COVID-19 / virology
  • Cell Surface Display Techniques / methods
  • Humans
  • Immunoglobulin Heavy Chains / immunology*
  • Immunoglobulin Variable Region / immunology*
  • Peptide Library*
  • SARS-CoV-2 / immunology*
  • SARS-CoV-2 / isolation & purification
  • Sequence Homology

Substances

  • Antibodies, Neutralizing
  • Antibodies, Viral
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Variable Region
  • Peptide Library