A sensitive immunoassay for porcine interferon-alpha

Vet Immunol Immunopathol. 1992 Jan 31;30(4):319-27. doi: 10.1016/0165-2427(92)90102-v.

Abstract

Two murine monoclonal antibodies (mAbs) directed against different epitopes on recombinant porcine interferon-alpha (IFN-alpha) were selected and used to construct a two-site ELISA. This ELISA, when performed in a one-step version, detected about 0.5 units ml-1 of IFN-alpha and showed similar sensitivity but better precision than a cytopathic effect inhibition bioassay. Estimates of IFN-alpha in tissue culture medium by the two assays correlated well. In contrast, one or several factors in porcine serum reduced the sensitivity of the ELISA. Measurements of IFN-alpha in porcine serum was, however, possible in a two-step version of the ELISA, with a sensitivity of about 1 unit IFN-alpha ml-1. Results of ELISA and bioassay agreed, except that the ELISA possibly produced false positive results in two out of a total of 91 sera negative in the bioassay. In addition, one of 23 sera positive in the bioassay was negative in the ELISA.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / biosynthesis
  • Antibodies, Monoclonal / immunology
  • Biological Assay
  • Culture Media
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Epitopes / immunology
  • Interferon Type I / immunology
  • Interferon-alpha / blood*
  • Mice
  • Recombinant Proteins
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Swine

Substances

  • Antibodies, Monoclonal
  • Culture Media
  • Epitopes
  • Interferon Type I
  • Interferon-alpha
  • Recombinant Proteins