Therapeutic human antibodies derived from PCR amplification of B-cell variable regions

Immunol Rev. 1992 Dec:130:69-85. doi: 10.1111/j.1600-065x.1992.tb01521.x.

Abstract

Despite advances in the in vitro immunization of human B cells (Borrebaeck et al. 1988) and the development of immunodeficient mice (McCune et al. 1988) for the reconstitution of the human immune system ex vivo, immortalization of antigen-specific human B cells remains the limiting step in the generation of human monoclonal antibodies. Typically this is performed with the aid of Epstein-Barr virus transformation followed by subcloning, confirmation of antigen binding and hybridization of the B lymphoblasts to a suitable fusion partner such as GLI-H7. This general approach is effective and widely used; however, it is time-consuming with erratic results. These were the immediate reasons we and others devised methods to directly obtain the variable regions from small numbers of human B cells (Larrick et al. 1987). The success of the PCR-based approach is illustrated above. In the present studies we successfully captured and stably produced antibodies from the V regions of two potent human anti-tetanus antibodies secreted by heteromyelomas that were too unstable for scale-up production. Although further preclinical evaluation of these antibodies is in progress, results to date indicate that the recombinant antibodies produced in myeloma-based cell lines or CHO cells are equivalent in binding specificity and activity to the native heteromyeloma-derived antibodies. Recent studies from this laboratory indicate that effective anti-tetanus protection will require a cocktail of anti-tetanus antibodies. Details of this work will be the subject of a future communication (Lang et al., in preparation).

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Bacterial / biosynthesis*
  • Antibodies, Bacterial / genetics
  • Antibodies, Bacterial / therapeutic use
  • Antibodies, Monoclonal / biosynthesis*
  • Antibodies, Monoclonal / genetics
  • Antibodies, Monoclonal / immunology
  • Antibodies, Monoclonal / therapeutic use
  • Base Sequence
  • CHO Cells
  • Cell Line, Transformed
  • Cricetinae
  • DNA / genetics
  • Enzyme-Linked Immunosorbent Assay
  • Genes, Immunoglobulin*
  • Humans
  • Hybridomas / immunology
  • Immunoglobulin Heavy Chains / genetics
  • Immunoglobulin Heavy Chains / immunology
  • Immunoglobulin Variable Region / genetics*
  • Immunoglobulin Variable Region / immunology
  • Mice
  • Molecular Sequence Data
  • Polymerase Chain Reaction*
  • Recombinant Fusion Proteins / biosynthesis*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / therapeutic use
  • Tetanus Toxoid / immunology*
  • Transfection

Substances

  • Antibodies, Bacterial
  • Antibodies, Monoclonal
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Variable Region
  • Recombinant Fusion Proteins
  • Tetanus Toxoid
  • DNA

Associated data

  • GENBANK/S42422
  • GENBANK/S42424
  • GENBANK/S42425
  • GENBANK/S42426
  • GENBANK/S55287
  • GENBANK/S55289
  • GENBANK/S55290
  • GENBANK/S55292
  • GENBANK/X62129
  • GENBANK/X62130