Schirrmann Thomas, Hust Michael
Department of Biotechnology, Institute of Biochemistry and Biotechnology, Technische Universität Braunschweig, Braunschweig, Germany.
Methods Mol Biol. 2010;651:177-209. doi: 10.1007/978-1-60761-786-0_11.
Recombinant antibodies as therapeutics offer new opportunities for the treatment of many tumor diseases. To date, 18 antibody-based drugs are approved for cancer treatment and hundreds of anti-tumor antibodies are under development. The first clinically approved antibodies were of murine origin or human-mouse chimeric. However, since murine antibody domains are immunogenic in human patients and could result in human anti-mouse antibody (HAMA) responses, currently mainly humanized and fully human antibodies are developed for therapeutic applications.Here, in vitro antibody selection technologies directly allow the selection of human antibodies and the corresponding genes from human antibody gene libraries. Antibody phage display is the most common way to generate human antibodies and has already yielded thousands of recombinant antibodies for research, diagnostics and therapy. Here, we describe methods for the construction of human scFv gene libraries and the antibody selection.
重组抗体作为治疗药物为许多肿瘤疾病的治疗提供了新的机遇。迄今为止,已有18种基于抗体的药物被批准用于癌症治疗,还有数百种抗肿瘤抗体正在研发中。首个获得临床批准的抗体来源于小鼠或人-鼠嵌合体。然而,由于小鼠抗体结构域在人类患者中具有免疫原性,可能会引发人抗小鼠抗体(HAMA)反应,目前主要开发人源化和全人源抗体用于治疗应用。在此,体外抗体筛选技术可直接从人抗体基因文库中筛选出人抗体及相应基因。抗体噬菌体展示是产生人抗体最常用的方法,已产生了数千种用于研究、诊断和治疗的重组抗体。在此,我们描述了构建人单链抗体基因文库和抗体筛选的方法。