Nahary Limor, Benhar Itai
Tel-Aviv University, Ramat Aviv, Israel.
Methods Mol Biol. 2009;525:61-80, xiv. doi: 10.1007/978-1-59745-554-1_3.
Antibody libraries came into existence 15 years ago when the accumulating sequence data of immunoglobulin genes and the advent of the PCR technology made it possible to clone antibody gene repertoires. Phage display (most common) and additional display and screening technologies were applied to pan out desired binding specificities from antibody libraries. "Synthetic" or "semi-synthetic" libraries are from naïve, non-immunized source and considered to be a source for many different targets, including self-antigens. We describe here how to construct a large human synthetic single-chain Fv (scFv) antibody library displayed on phages, where in vivo-formed complementarity-determining regions (CDRs) are shuffled combinatorially onto germline-derived human variable-region frameworks.
抗体文库诞生于15年前,当时免疫球蛋白基因的序列数据不断积累,PCR技术的出现使得克隆抗体基因库成为可能。噬菌体展示(最常用)以及其他展示和筛选技术被用于从抗体文库中淘选出所需的结合特异性。“合成”或“半合成”文库来自未经免疫的天然来源,被认为是许多不同靶点(包括自身抗原)的来源。我们在此描述如何构建一个展示在噬菌体上的大型人源合成单链Fv(scFv)抗体文库,其中体内形成的互补决定区(CDR)被组合重排到种系衍生的人可变区框架上。