Biologics Research, Merck Research Laboratories, Merck & Co., Inc., Palo Alto, CA 94304, USA.
Protein Eng Des Sel. 2011 Sep;24(9):711-9. doi: 10.1093/protein/gzr034. Epub 2011 Jul 12.
We created a cross-species display system that allows the display of the same antibody libraries on both prokaryotic phage and eukaryotic yeast without the need for molecular cloning. Using this cross-display system, a large, diverse library can be constructed once and subsequently used for display and selection in both phage and yeast systems. In this article, we performed the parallel phage and yeast selection of an antibody maturation library using this cross-display platform. This parallel selection allowed us to isolate more unique hits than single-species selection, with 162 unique clones from phage and 107 unique clones from yeast. In addition, we were able to shuttle yeast hits back to Escherichia coli cells for affinity characterization at a higher throughput.
我们创建了一个跨物种展示系统，可以在原核噬菌体和真核酵母上展示相同的抗体文库，而无需分子克隆。使用这种跨展示系统，可以一次构建一个大型的、多样化的文库，然后在噬菌体和酵母系统中进行展示和选择。在本文中，我们使用这种跨展示平台对一个抗体成熟文库进行了噬菌体和酵母的平行选择。这种平行选择使我们能够分离出比单一物种选择更多的独特命中物，从噬菌体中分离出 162 个独特克隆，从酵母中分离出 107 个独特克隆。此外，我们还能够将酵母命中物转移回大肠杆菌细胞，以更高的通量进行亲和力表征。
