Simple workflow to repurpose SARS-CoV-2 swab/serum samples for the isolation of cost-effective antibody/antigens for proteotyping applications and diagnosis

Anal Bioanal Chem. 2021 Dec;413(29):7251-7263. doi: 10.1007/s00216-021-03654-4. Epub 2021 Oct 8.

Abstract

Supply shortage for the development and production of preventive, therapeutic, and diagnosis tools during the COVID-19 pandemic is an important issue affecting the wealthy and poor nations alike. Antibodies and antigens are especially needed for the production of immunological-based testing tools such as point-of-care tests. Here, we propose a simple and quick magnetic nanoparticle (MNP)-based separation/isolation approach for the repurposing of infected human samples to produce specific antibodies and antigen cocktails. Initially, an antibody cocktail was purified from serums via precipitation and immunoaffinity chromatography. Purified antibodies were conjugated onto MNPs and used as an affinity matrix to separate antigens. The characterization process was performed by ELISA, SDS-PAGE, electrochemistry, isothermal titration calorimetry, and LC-Q-TOF-MS/MS analyses. The MNP-separated peptides can be used for mass spectrometry-based as well as paper-based lateral flow assay diagnostic. The exploitation of the current workflow for the development of efficient diagnostic tools, specific treatments, and fundamental research can significantly impact the present or eventual pandemic. This workflow can be considered as a two birds, one stone-like strategy.

Keywords: Antigen/antibody purification; Lateral flow assay; Magnetic nanoparticle (MNP)–based separation; Mass spectrometry; SARS-CoV-2.

MeSH terms

  • Antibodies, Viral / blood
  • Antibodies, Viral / isolation & purification*
  • Antigens, Viral / blood
  • Antigens, Viral / isolation & purification*
  • COVID-19 / diagnosis*
  • COVID-19 / virology
  • Calorimetry
  • Cost-Benefit Analysis*
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Immunoassay / economics*
  • SARS-CoV-2 / immunology
  • SARS-CoV-2 / isolation & purification*
  • Specimen Handling
  • Tandem Mass Spectrometry
  • Viremia / blood
  • Viremia / virology*
  • Workflow

Substances

  • Antibodies, Viral
  • Antigens, Viral