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14Selection of Recombinant AntibodiesFrom Antibody Gene LibrariesMichael Hust, Stefan Dübel, and Thomas SchirrmannSummaryAfter the sequencing of the human genome is completed, the research focus shifts toward theanalysis of gene products. The human genome encodes more than 30,000 genes. Owing to alternativemRNA splicing and posttranslational modifications, for example, glycosylation, phoshorylation,and so on, the number of different proteins of human proteome is supposed to easilyexceed 90,000. Antibodies are key detection reagents for the “postgenomic” analysis of these proteins.Any systematic investigation of the human proteome requires high throughput methods forantibody generation. In vitro selection systems utilizing recombinant antibody repertoires offerthis capability and capacity. The most commonly used contemporary in vitro selection system isantibody phage display, which has already yielded thousands of useful antibodies for therapy,research, and diagnostics. Herein, methods are described for the selection of recombinant antibodyfragments from naive antibody gene libraries.Key Words: Antibody engineering; panning; phage display; scFv; antibody gene libraries;filamentous phage.1. IntroductionThe production of polyclonal antibodies by immunization of animals isestablished for more than a century. The first antibody serum was directedagainst diphtheria and produced in horses (1). Hybridoma technology was thenext milestone, allowing the production of monoclonal antibodies by fusion ofan antibody producing spleen B-cell with an immortal myeloma cell (2).However, hybridoma technology has some limitations like a potential geneticinstability of the aneuploid cell lines and most of all its inability to produceantibodies against toxic or highly conserved antigens (3). When repeatedlyadministered in therapy, murine hybridoma antibodies induce a human antimouseantibody response caused by murine antibodies (4). This problem can beFrom: Methods in Molecular Biology, vol. 408: Gene Function AnalysisEdited by: M. Ochs © Humana Press Inc., Totowa, NJ243