Characterization of purified protein derivative of tuberculin by use of monoclonal antibodies: isolation of a delayed-type hypersensitivity reactive component from M. tuberculosis culture filtrate

Scand J Immunol. 1994 Sep;40(3):345-9. doi: 10.1111/j.1365-3083.1994.tb03471.x.

Abstract

Nine monoclonal antibodies were raised against purified protein derivative (PPD) of tuberculin in mice previously treated with Bacilli Calmette Guérin (BCG). The antibodies also reacted with a culture filtrate from Mycobacterium tuberculosis strain H37Rv. In immunoblotting after SDS-PAGE the reaction with PPD was seen as a diffuse smear, whereas ammonium sulphate-precipitated proteins from H37Rv gave well-defined bands ranging from 10 to 65 kDa. Enzyme immunoassay showed that both PPD and H37Rv antigens were able to inhibit binding of the antibodies to PPD coated microtitre wells, suggesting that the antibodies reacted with continuous epitopes. A 12 kDa protein purified by immunoaffinity chromatography from H37Rv antigens was tested intradermally in M. tuberculosis MNC3 sensitized guinea pigs and gave a delayed type hypersensitivity reaction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology
  • Electrophoresis, Polyacrylamide Gel
  • Guinea Pigs
  • Humans
  • Hypersensitivity, Delayed / immunology*
  • Immunoblotting
  • Immunoenzyme Techniques
  • Mice
  • Mice, Inbred C57BL
  • Mycobacterium tuberculosis / immunology*
  • Rosaniline Dyes
  • Tuberculin / immunology*

Substances

  • Antibodies, Monoclonal
  • Rosaniline Dyes
  • Tuberculin
  • coomassie Brilliant Blue