Figure 2.

Combinatorial mining of human antibody repertoires. Polyclonal or oligoclonal mixtures of human B cells (A) are used in bulk to isolate mRNA and amplify cDNA of heavy and light chains (B). (C) Heavy and light chain cDNAs are cloned into display vectors that afford antibody libraries with a physical linkage of genotype (cDNA) and phenotype (protein). Shown as examples are filamentous phage that encode and display Fab (top) and virus-infected mammalian cells that express and display scFv (bottom). Note that heavy and light chains are randomly combined. (D) Antibody libraries are then selected by several rounds of panning on immobilized antigen (top) or screened by a single round of FACS using fluorescently labeled antigen (bottom). (See text for details). The physical linkage of genotype and phenotype allows the enrichment and amplification of displayed antibodies. (E) Human mAbs from antibody libraries are manufactured the same way as human mAbs from monoclonal B cells and B-cell lines.