Selection and characterization of scFv antibodies against the Sin Nombre hantavirus nucleocapsid protein

J Immunol Methods. 2007 Apr 10;321(1-2):60-9. doi: 10.1016/j.jim.2007.01.011. Epub 2007 Feb 8.

Abstract

Rodent-borne hantaviruses cause hemorrhagic fever with renal syndrome (HFRS) in the old world and hantavirus cardio-pulmonary syndrome (HCPS) in the new. Most cases of HCPS in North America are caused by Sin Nombre Virus (SNV). Current viral detection technologies depend upon the identification of anti-viral antibodies in patient serum. Detection of viral antigen may facilitate earlier detection of the pathogen. We describe here the characterization of two single-chain Fv antibodies (scFvs), selected from a large naïve phage antibody library, which are capable of identifying the Sin Nombre Virus nucleocapsid protein (SNV-N), with no cross reactivity with the nucleocapsid protein from other hantaviruses. The utility of such selected scFvs was increased by the creation of an scFv-alkaline phosphatase fusion protein which was able to directly detect virally produced material without the need for additional reagents.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaline Phosphatase / genetics
  • Alkaline Phosphatase / immunology
  • Animals
  • Antibodies, Viral / genetics
  • Antibodies, Viral / immunology*
  • Antibody Affinity
  • Antibody Specificity
  • Chlorocebus aethiops
  • Cloning, Molecular
  • Cross Reactions
  • Enzyme-Linked Immunosorbent Assay
  • Hantavirus Pulmonary Syndrome / diagnosis
  • Hantavirus Pulmonary Syndrome / immunology*
  • Immunoglobulin Variable Region / genetics
  • Immunoglobulin Variable Region / immunology*
  • Nucleocapsid Proteins / analysis
  • Nucleocapsid Proteins / genetics
  • Nucleocapsid Proteins / immunology*
  • Peptide Library
  • Recombinant Fusion Proteins / immunology
  • Recombinant Proteins / immunology
  • Sin Nombre virus / genetics
  • Sin Nombre virus / immunology*
  • Vero Cells

Substances

  • Antibodies, Viral
  • Immunoglobulin Variable Region
  • Nucleocapsid Proteins
  • Peptide Library
  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • Alkaline Phosphatase