From cells to genes: how to make antibodies useful in human diagnosis and therapy

Monoclonal antibodies (mAb) have great potential value for in vivo diagnosis and therapy in humans. Their antigenic nature is however responsible for severe side effects and successful applications in the clinic have prove to be more limited than was originally hoped. This review summarize both cell biology and molecular biology approaches developed in order to overcome these limitations. Improving methodologies to immortalize cell lines producing human mAbs are reported with a particular attention to the techniques aimed at rescuing B cells expressing high-affinity human antibodies. A major part of this review is devoted to the protein engineering work. Genetic manipulation of mouse monoclonals to produce humanized antibodies and preparation of bacteriophage libraries displaying Ig repertoires are examined. The possibility to extend these approaches to the production of in vitro repertoires and to obviate the in vivo immunization step is also discussed.