Isolation and identification of an scFv antibody against nucleocapsid protein of SARS-CoV

Microbes Infect. 2007 Jul;9(8):1026-33. doi: 10.1016/j.micinf.2007.04.008. Epub 2007 Apr 21.

Abstract

To develop reagents for early diagnosis and therapeutic drugs against SARS-associated coronavirus (SARS-CoV), a large (3 x 10(9)) immunized human antibody library was constructed from peripheral blood mononuclear cells from six SARS convalescent patients. A single chain variable fragment antibody (N18) with high affinity against N protein of SARS-CoV was isolated. Sequence analysis revealed that the VL gene was composed of VL3h (V lambda subgroup) and JL2 regions and the VH gene was composed of VH1-69 (VH1 subgroup), D2-15, D3-22 and JH6 regions. Soluble N18 antibody was expressed in Escherichia coli HB2151, purified by Ni-NTA affinity chromatography and verified by SDS-PAGE and Western blot. The potential application for early diagnosis was evaluated using N protein capture ELISA in which N18 antibody demonstrated high sensitive activity in detecting N protein of SARS-CoV. Finally, the potential usefulness of the N18 antibody in prophylaxis, vaccine design and therapy of SARS is discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antibodies, Viral / immunology*
  • Antibody Specificity*
  • Base Sequence
  • Coronavirus Nucleocapsid Proteins
  • Humans
  • Immunoglobulin Fragments / chemistry
  • Immunoglobulin Fragments / genetics
  • Immunoglobulin Fragments / immunology*
  • Molecular Sequence Data
  • Nucleocapsid Proteins / immunology*
  • Peptide Library*
  • Sequence Analysis, DNA
  • Severe acute respiratory syndrome-related coronavirus / immunology*

Substances

  • Antibodies, Viral
  • Coronavirus Nucleocapsid Proteins
  • Immunoglobulin Fragments
  • Nucleocapsid Proteins
  • Peptide Library