Preparation and characterization of monoclonal antibodies against S1 domain at N-terminal residues 249 to 667 of SARS-associated coronavirus S1 protein

Di Yi Jun Yi Da Xue Xue Bao. 2004 Jan;24(1):1-6.

Abstract

Objective: To prepare and characterize monoclonal antibodies (mAbs) against S1 protein of severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV).

Methods: 6-His-tagged recombinant fragment at N-terminal residues 249 to 667 of SARS-CoV S1 protein including S-protein receptor-binding domain was expressed in E.coli. The immunogenicity of this S1 domain was identified and used to immunize BALB/c mice for the production of hybridomas. The identification of the mAbs against this S1 domain was performed using indirect enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay (IFA) and Western blotting, respectively.

Results: Three hybridomas producing mAbs specific to the S1 domain was obtained, with a relative molecular mass of 48,500. None of the 3 mAbs were reactive with human coronaviruses 229E and OC43. Two of the mAbs were IgG2a isotype, and the other was IgG1.

Conclusions: This is the first report of mAbs produced against S-protein receptor-binding domain of SARS-CoV. The 3 S1-specific mAbs may be useful for further study of the function of the S protein and for diagnosis of SARS-CoV infection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / biosynthesis
  • Antibodies, Monoclonal / immunology*
  • Blotting, Western
  • Cell Line
  • Female
  • Fluorescent Antibody Technique, Indirect
  • Humans
  • Membrane Glycoproteins / immunology*
  • Membrane Glycoproteins / physiology
  • Mice
  • Mice, Inbred BALB C
  • Peptide Fragments / immunology*
  • Severe Acute Respiratory Syndrome / diagnosis
  • Severe acute respiratory syndrome-related coronavirus / chemistry*
  • Spike Glycoprotein, Coronavirus
  • Viral Envelope Proteins / immunology*
  • Viral Envelope Proteins / physiology

Substances

  • Antibodies, Monoclonal
  • Membrane Glycoproteins
  • Peptide Fragments
  • Spike Glycoprotein, Coronavirus
  • Viral Envelope Proteins
  • spike glycoprotein, SARS-CoV
  • spike protein, mouse hepatitis virus