Tabata Noriko, Sakuma Yuko, Honda Yumiko, Doi Nobuhide, Takashima Hideaki, Miyamoto-Sato Etsuko, Yanagawa Hiroshi
Department of Biosciences and Informatics, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-8522, Japan.
Nucleic Acids Res. 2009 May;37(8):e64. doi: 10.1093/nar/gkp184. Epub 2009 Mar 30.
In vitro antibody-display technologies are powerful approaches for isolating monoclonal antibodies from recombinant antibody libraries. However, these display techniques require several rounds of affinity selection which is time-consuming. Here, we combined mRNA display with a microfluidic system for in vitro selection and evolution of antibodies and achieved ultrahigh enrichment efficiency of 10(6)- to 10(8)-fold per round. After only one or two rounds of selection, antibodies with high affinity and specificity were obtained from naive and randomized single-chain Fv libraries of approximately 10(12) molecules. Furthermore, we confirmed that not only protein-protein (antigen-antibody) interactions, but also protein-DNA and protein-drug interactions were selected with ultrahigh efficiencies. This method will facilitate high-throughput preparation of antibodies and identification of protein interactions in proteomic and therapeutic fields.
体外抗体展示技术是从重组抗体文库中分离单克隆抗体的强大方法。然而，这些展示技术需要进行几轮亲和力选择，这很耗时。在此，我们将mRNA展示与微流控系统相结合用于抗体的体外选择和进化，每轮实现了10⁶至10⁸倍的超高富集效率。仅经过一轮或两轮选择，就从约10¹²个分子的天然和随机单链Fv文库中获得了具有高亲和力和特异性的抗体。此外，我们证实不仅蛋白质-蛋白质（抗原-抗体）相互作用，而且蛋白质-DNA和蛋白质-药物相互作用也能以超高效率被筛选出来。该方法将有助于在蛋白质组学和治疗领域高通量制备抗体以及鉴定蛋白质相互作用。
