Generation of human monoclonal antibodies against HIV-1 proteins; electrofusion and Epstein-Barr virus transformation for peripheral blood lymphocyte immortalization

AIDS Res Hum Retroviruses. 1994 Apr;10(4):359-69. doi: 10.1089/aid.1994.10.359.


Electrofusion and EBV transformation were studied by immortalizing human PBLs from blood of HIV-1-positive volunteers. A panel of 33 cell lines producing human monoclonal antibodies (Hu-MAbs) against HIV-1 was established by cell fusion or EBV transformation. For the first fusion experiments the source of B lymphocytes was peripheral blood of HIV-1-infected donors in CDC stages II or III with CD4 cell counts higher than 500/mm3. Later on, from these patients only, those with high anti-HIV titers were chosen as blood donors. By that means the yield of stable specific hybridomas was increased twofold. In our experiments electrofusion turned out to be a more efficient immortalization method than EBV transformation, due to a high and constant immortalization rate. The hybridomas were stable after intensive subcloning and could be cultivated over a period of 8 months without loss in monoclonal antibody production. Immunoglobulin class, subtype, reactivity against HIV-1 proteins, Western blot patterns, immunofluorescence, and epitopes were characterized. The subtype of all antibodies was IgG1 or IgG3. The light chain was predominantly kappa. All antibodies showed reactivity against HIV-1 envelope or core protein. All hybridomas were stable and suited for mass production. Several Hu-MAbs are becoming an important tool in the field of diagnosis, research, and immunotherapy.

MeSH terms

  • Antibodies, Monoclonal / biosynthesis*
  • Antibodies, Monoclonal / chemistry
  • B-Lymphocytes / immunology
  • Cell Fusion
  • Cell Line
  • Cell Transformation, Viral
  • Epitopes
  • HIV Antibodies / biosynthesis*
  • HIV Antibodies / chemistry
  • HIV Envelope Protein gp120 / immunology
  • HIV Infections / immunology
  • HIV-1 / immunology*
  • Herpesvirus 4, Human
  • Humans
  • Hybridomas / immunology
  • Immunochemistry
  • In Vitro Techniques
  • Neutralization Tests


  • Antibodies, Monoclonal
  • Epitopes
  • HIV Antibodies
  • HIV Envelope Protein gp120