Kenrick Sophia, Rice Jeffrey, Daugherty Patrick
University of California, Santa Barbara, California, USA.
Curr Protoc Cytom. 2007 Oct;Chapter 4:Unit4.6. doi: 10.1002/0471142956.cy0406s42.
The protocols herein detail methods for isolating binding peptides from a combinatorial library displayed on the surface of bacterial cells. These methods are appropriate for a variety of display scaffolds and a large range of library sizes, up to approximately 5 x 10(9) or more. Instructions have been provided for isolating peptides that bind to both proteins and non-protein targets, such as whole cells or inorganic particles. Qualitative analysis by flow cytometry can be exploited for bacterial libraries to characterize a displayed peptide's binding properties with a target of interest, and sorting conditions can be tuned to maximize binding affinity.
本文中的方案详细介绍了从展示在细菌细胞表面的组合文库中分离结合肽的方法。这些方法适用于多种展示支架和各种文库规模，最大可达约5×10⁹或更多。已提供了分离与蛋白质和非蛋白质靶标（如全细胞或无机颗粒）结合的肽的说明。对于细菌文库，可利用流式细胞术进行定性分析，以表征展示肽与感兴趣靶标的结合特性，并且可以调整分选条件以最大化结合亲和力。
