Protocol for evaluating humoral immune responses in mice following SARS-CoV-2 vaccination

STAR Protoc. 2025 Mar 21;6(1):103548. doi: 10.1016/j.xpro.2024.103548. Epub 2025 Jan 15.

Abstract

Binding and neutralizing antibodies are critical indicators of protection against viral pathogens and are essential for assessing the immunogenicity and efficacy of a vaccine. Here, we present a protocol comprising two assays for measuring the spike-specific binding and neutralizing antibodies in mouse plasma following severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccination. We describe steps for determining binding antibody titers using enzyme-linked immunosorbent assay (ELISA) and assessing neutralizing antibody titers through a pseudovirus neutralization assay. For complete details on the use and execution of this protocol, please refer to Jingyou Yu et al.1.

Keywords: cell biology; cell-based assays; immunology; microbiology; molecular biology.

MeSH terms

  • Animals
  • Antibodies, Neutralizing / blood
  • Antibodies, Neutralizing / immunology
  • Antibodies, Viral* / blood
  • Antibodies, Viral* / immunology
  • COVID-19 Vaccines* / immunology
  • COVID-19* / immunology
  • COVID-19* / prevention & control
  • Enzyme-Linked Immunosorbent Assay / methods
  • Female
  • Humans
  • Immunity, Humoral* / immunology
  • Mice
  • Neutralization Tests / methods
  • SARS-CoV-2* / immunology
  • Spike Glycoprotein, Coronavirus / immunology
  • Vaccination

Substances

  • COVID-19 Vaccines
  • Antibodies, Neutralizing
  • Antibodies, Viral
  • Spike Glycoprotein, Coronavirus