Development of Whole-Porcine Monoclonal Antibodies with Potent Neutralization Activity against Classical Swine Fever Virus from Single B Cells

ACS Synth Biol. 2019 May 17;8(5):989-1000. doi: 10.1021/acssynbio.8b00365. Epub 2019 Apr 12.

Abstract

Classical swine fever (CSF) is a highly contagious swine disease that causes devastating economic losses. However, there are few efficacious therapeutic antibodies against the CSF virus (CSFV). Accordingly, we isolated two whole-porcine anti-CSFV neutralizing antibodies (NAbs) directly from single B cells sorted using the conserved linear epitope of the CSFV E2 protein and goat anti-pig IgG. These mAbs, termed HK24 and HK44, can bind to the E2 protein by recognizing sites within the conserved linear epitope of E2. In addition, these two mAbs can detect virus infection with high specificity and possess potent neutralizing activity. HK24 and HK44 protect PK-15 cells from CSFV infections in vitro with potent IC50 values of 9.3 and 0.62 μg/mL, respectively. We anticipate that these antibodies can be used as diagnostic and antiviral agents for CSFV and that the method we describe here will accelerate the production of therapeutic antibodies for other viruses.

Keywords: classical swine fever; epitope of E2 protein; single B cells; whole-porcine anti-CSFV neutralizing antibodies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal / genetics*
  • Antibodies, Monoclonal / metabolism
  • Antibodies, Viral / genetics*
  • Antibodies, Viral / metabolism
  • Antigen-Antibody Reactions
  • B-Lymphocytes / cytology
  • B-Lymphocytes / metabolism*
  • Cell Line
  • Classical Swine Fever Virus / immunology*
  • Classical Swine Fever Virus / metabolism
  • Epitopes / chemistry
  • Epitopes / immunology
  • Epitopes / metabolism
  • HEK293 Cells
  • Humans
  • Immunoglobulin Heavy Chains / genetics
  • Immunoglobulin Light Chains / genetics
  • Plasmids / genetics
  • Plasmids / metabolism
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Single-Cell Analysis
  • Swine
  • Viral Envelope Proteins / genetics
  • Viral Envelope Proteins / immunology*
  • Viral Envelope Proteins / metabolism

Substances

  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Epitopes
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Light Chains
  • Recombinant Proteins
  • Viral Envelope Proteins
  • glycoprotein E2, classical swine fever virus