The Shmunis School of Biomedicine and Cancer Research, The George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, Israel.
Ukko LTD, Rehovot, Israel.
Methods Mol Biol. 2023;2702:347-372. doi: 10.1007/978-1-0716-3381-6_18.
Antibody libraries came into existence 30 years ago when the accumulating sequence data of immunoglobulin genes and the advent of 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. As other antibody discovery tools, phage display is not an off-the-shelf technology and not offered as a kit but rather requires experience and expertise for making it indeed very useful.Next-generation sequencing (NGS) coupled with bioinformatics is a powerful tool for analyzing large amount of DNA sequence output of the panning. Here, we demonstrate how NGS analysis of phage biopanning (phage-Seq) of complex antibody libraries can facilitate the antibody discovery process and provide insights regarding the biopanning process (see Fig. 1).
抗体库诞生于 30 年前，当时免疫球蛋白基因的序列数据不断积累，PCR 技术也问世了，这使得克隆抗体基因库成为可能。噬菌体展示（最常见）和其他展示和筛选技术被应用于从抗体库中筛选出所需的结合特异性。与其他抗体发现工具一样，噬菌体展示不是一种现成的技术，也不是作为试剂盒提供的，而是需要经验和专业知识才能真正发挥其作用。下一代测序（NGS）与生物信息学相结合，是分析淘选过程中大量 DNA 序列输出的强大工具。在这里，我们展示了如何通过对复杂抗体库的噬菌体淘选（噬菌体-Seq）进行 NGS 分析，来促进抗体发现过程，并提供有关淘选过程的见解（见图 1）。
