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Baculovirus-Free SARS-CoV-2 Virus-like Particle Production in Insect Cells for Rapid Neutralization Assessment.

Marcel Jaron1, Michael Lehky2, Marta Zarà3

  • 1Department of Biotechnology, Technische Universität Braunschweig, Spielmannstraße 7, 38106 Braunschweig, Germany.

Viruses
|October 27, 2022
PubMed
Summary
This summary is machine-generated.

We developed a rapid method to produce SARS-CoV-2 virus-like particles (VLPs) in insect cells. These VLPs efficiently evaluate the effectiveness of antibodies and sera from vaccinated individuals against the virus.

Keywords:
SARS-CoV-2antibodiescellular assayexpression vectorinsect cellsvirus-like particles (VLPs)

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Area of Science:

  • Virology
  • Immunology
  • Biotechnology

Background:

  • Virus-like particles (VLPs) mimic authentic viruses but lack genetic material, making them valuable research tools.
  • Current methods for producing SARS-CoV-2 VLPs in insect cells often rely on baculovirus systems, which can be time-consuming.

Purpose of the Study:

  • To establish a fast and efficient method for producing SARS-CoV-2 VLPs in insect cells.
  • To utilize these VLPs for assessing the inhibitory potential of monoclonal antibodies and sera from vaccinated individuals.

Main Methods:

  • Direct plasmid transfection was used to co-express SARS-CoV-2 envelope, membrane, and spike proteins, leading to VLP self-assembly.
  • Nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM) confirmed VLP characteristics (~145 nm diameter, 'Corona' appearance).
  • Membrane protein fused to GFP enabled direct quantification of binding inhibition to ACE2 via flow cytometry.

Main Results:

  • Optimized expression plasmids and vector ratios yielded well-formed SARS-CoV-2 VLPs.
  • The GFP fusion facilitated straightforward assessment of antibody and serum inhibition of VLP-ACE2 binding.
  • The assay obviated the need for VLP purification or secondary fluorescent antibody labeling.

Conclusions:

  • A novel, rapid, and powerful method for producing SARS-CoV-2 VLPs in insect cells was developed, bypassing traditional baculovirus approaches.
  • The produced VLPs are suitable for evaluating the efficacy of therapeutic antibodies and immune sera.
  • The developed flow cytometry assay is efficient and does not require VLP purification or secondary labeling.