A high-throughput SARS-CoV-2 pseudovirus multiplex neutralization assay

STAR Protoc. 2022 Dec 16;3(4):101835. doi: 10.1016/j.xpro.2022.101835. Epub 2022 Oct 19.


Evaluating the neutralizing antibody titer following SARS-CoV-2 vaccination is essential in defining correlates of protection. We describe an assay that uses single-cycle vesicular stomatitis virus (VSV) pseudoviruses linking a fluorophore with a spike (S) from a variant of concern (VOC). Using two fluorophores linked to two VOC S, respectively, allows us to determine the neutralization titer against two VOCs in a single run. This is a generalizable approach that saves time, samples, and run-to-run variability. For complete details on the use and execution of this protocol, please refer to Sievers et al. (2022).1.

Keywords: Antibody; Cell-based Assays; High Throughput Screening; Microbiology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Viral
  • COVID-19 Vaccines
  • COVID-19*
  • Humans
  • Neutralization Tests / methods
  • SARS-CoV-2*


  • COVID-19 Vaccines
  • Antibodies, Viral