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A simple protein-based surrogate neutralization assay for SARS-CoV-2.

Kento T Abe1,2, Zhijie Li1, Reuben Samson1,2

  • 1Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada.

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|September 2, 2020
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Summary

A new protein-based assay can detect SARS-CoV-2 antibodies that block virus entry, correlating with neutralization. This offers a safer, efficient alternative for assessing immune response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

Keywords:
Adaptive immunityAntigenImmunoglobulinsInfectious disease

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Area of Science:

  • Immunology
  • Virology
  • Biotechnology

Background:

  • Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection elicits a humoral immune response, but its duration and clinical correlation remain unclear.
  • Identifying immune correlates is crucial for guiding public health decisions on treatments, vaccines, and convalescent plasma therapy.
  • Current methods for detecting neutralizing antibodies against SARS-CoV-2 are often complex and require cell-based assays.

Purpose of the Study:

  • To develop and validate a safe and efficient protein-based assay for detecting SARS-CoV-2 neutralizing antibodies.
  • To serve as a surrogate neutralization assay that directly compares with ELISA-based antibody detection.
  • To correlate assay results with established viral-based neutralization assays.

Main Methods:

  • Developed a protein-based assay to detect antibodies blocking the SARS-CoV-2 spike protein's receptor binding domain (RBD) interaction with ACE2.
  • Performed the assay in parallel with an ELISA for RBD-specific antibodies on the same platform.
  • Validated the assay against a plaque reduction neutralization test (PRNT) using live SARS-CoV-2 and a spike pseudotyped viral vector assay.

Main Results:

  • The protein-based assay successfully detected serum and plasma antibodies that inhibit the RBD-ACE2 interaction.
  • Assay results showed strong correlation with viral-based neutralization assays, including PRNT.
  • The assay provides a surrogate measure of neutralization on a platform compatible with standard ELISA.

Conclusions:

  • The developed protein-based assay is a safe and efficient surrogate for viral neutralization assays.
  • This assay can aid in characterizing the duration and clinical relevance of SARS-CoV-2 humoral immunity.
  • The assay facilitates direct comparison with antibody detection assays, supporting public health strategies.