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A bovine monoclonal antibody detecting a class I BoLA antigen

S J Kemp1, E M Tucker, A J Teale

  • 1International Laboratory for Research on Animal Diseases, Nairobi, Kenya.

Animal Genetics
|January 1, 1990
PubMed

Insights

Researchers developed a new bovine monoclonal antibody (mAb) called ILA70. This antibody specifically targets a polymorphic epitope on bovine class I major histocompatibility complex (MHC) molecules, aiding in cattle immune system research.

Area of Science:

  • Immunology
  • Veterinary Science
  • Genetics

Background:

  • Monoclonal antibodies (mAbs) are crucial tools for characterizing cell surface molecules.
  • The bovine major histocompatibility complex (MHC) plays a vital role in immune responses and disease resistance in cattle.
  • Developing specific mAbs for bovine MHC molecules facilitates detailed serological analysis.

Purpose of the Study:

  • To generate and characterize a novel bovine monoclonal antibody (mAb) for identifying bovine class I MHC molecules.
  • To assess the utility of the mAb ILA70 in serological typing and immune studies.
  • To establish a stable hybridoma for producing high-affinity antibodies.

Main Methods:

  • Production of mAb ILA70 through immunization of a calf with peripheral blood mononuclear cells (PBM) and subsequent heterohybridoma fusion.
  • Characterization of the target epitope using antibody binding inhibition and immunoprecipitation assays.
  • Evaluation of complement-fixing activity for cytotoxicity assays.
  • Preparation of ascitic fluid for enhanced antibody yield.
  • Subcloning of the antibody-producing heterohybridoma to ensure stability.

Main Results:

  • The mAb ILA70 was successfully generated and identified as recognizing a polymorphic epitope on a bovine class I MHC molecule.
  • The antibody exhibits complement-fixing properties, making it suitable for standard cytotoxicity assays.
  • Ascitic fluid preparation significantly increased antibody activity compared to tissue-culture supernatant.
  • The antibody-producing heterohybridoma demonstrated stability through multiple subcloning steps.

Conclusions:

  • The mAb ILA70 is a valuable tool for the precise serological analysis of bovine class I MHC products.
  • Heterohybridomas represent a promising source for generating highly specific and informative monoclonal antibodies for bovine MHC research.
  • This antibody can contribute to a deeper understanding of cattle immune genetics and disease management.

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