Figure 2. Generation of human monoclonal antibodies. (Phage display) Heavy and light chain cDNA isolated from human B-cells is used to generate a combinatorial library in which random heavy (H) and light chain (L) pairings are expressed on the surface of phage. These phage can then be screened for antigen binding by traditional techniques (e.g., ELISA). Since only the antigen binding region is used in the phage display process, the selected clone is then placed into an appropriate expression vector to produce a full antibody molecule. (Transgenics) Genetically manipulated mice have been produced with inactivated endogenous immuno- globulin genes, and with unrearranged human immunoglobulin gene segments introduced (90,91). These mice are then immunized with antigen, and hybridomas are produced by traditional routes. (See refs. 88, 89 for more technical information on these two methods and refs. 92, 93 for comparisons of these two methods).
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