Practice question

Hybridoma technology developed by Georges Köhler and César Milstein in 1975 and awarded Nobel 1984 created paradigm shift by immortalizing antibody-producing B lymphocytes through somatic cell fusion. Mice immunized with antigen develop germinal center reaction where B cells undergo VDJ recombination, somatic hypermutation and class switching to generate high affinity plasma cells. Splenic lymphocytes harvested shortly after boost are fused with HGPRT-deficient myeloma partner such as SP2/0-Ag14 or NS0 using polyethylene glycol 1500 that perturbs lipid bilayers promoting membrane fusion forming heterokaryons containing nuclei from both parents. Resulting hybrid cells combine immortal growth driven by myeloma oncogenes like c-myc deregulation with functional immunoglobulin heavy and light chain transcription from B cell. Clonal expansion and selection yields everlasting lines secreting monospecific antibody of defined isotype typically IgG1 kappa recognizing single epitope, known as monoclonal antibody. Such reagents revolutionized diagnostics, research tools like western blotting, and therapeutics by提供 unlimited standardized antibody with consistent affinity replacing heterogeneous polyclonal sera plagued by batch variation.