Hust Michael, Dübel Stefan, Schirrmann Thomas
Technical University Braunschweig, Germany.
Methods Mol Biol. 2007;408:243-55. doi: 10.1007/978-1-59745-547-3_14.
After the sequencing of the human genome is completed, the research focus shifts toward the analysis of gene products. The human genome encodes more than 30,000 genes. Owing to alternative mRNA splicing and posttranslational modifications, for example, glycosylation, phoshorylation, and so on, the number of different proteins of human proteome is supposed to easily exceed 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 for antibody generation. In vitro selection systems utilizing recombinant antibody repertoires offer this capability and capacity. The most commonly used contemporary in vitro selection system is antibody phage display, which has already yielded thousands of useful antibodies for therapy, research, and diagnostics. Herein, methods are described for the selection of recombinant antibody fragments from naive antibody gene libraries.
人类基因组测序完成后，研究重点转向基因产物的分析。人类基因组编码超过30000个基因。由于存在可变mRNA剪接以及翻译后修饰，例如糖基化、磷酸化等，人类蛋白质组中不同蛋白质的数量应该很容易超过90000种。抗体是对这些蛋白质进行“后基因组”分析的关键检测试剂。对人类蛋白质组进行任何系统性研究都需要高通量的抗体生成方法。利用重组抗体库的体外筛选系统具备这种能力。目前最常用的体外筛选系统是抗体噬菌体展示，它已经产生了数千种用于治疗、研究和诊断的有用抗体。本文描述了从天然抗体基因文库中筛选重组抗体片段的方法。
