Kay Brian K, Castagnoli Luisa
Argonne National Laboratory, Argonne, Illinois, USA.
Curr Protoc Cell Biol. 2003 Feb;Chapter 17:Unit 17.4. doi: 10.1002/0471143030.cb1704s17.
This unit describes the process and analysis of affinity selecting bacteriophage M13 from libraries displaying combinatorial peptides fused to either a minor or major capsid protein. Direct affinity selection uses target protein bound to a microtiter plate followed by purification of selected phage by ELISA. Alternatively, there is a bead-based affinity selection method. These methods allow one to readily isolate peptide ligands that bind to a protein target of interest and use the consensus sequence to search proteomic databases for putative interacting proteins.
本单元描述了从展示与次要或主要衣壳蛋白融合的组合肽的文库中亲和选择噬菌体M13的过程和分析。直接亲和选择使用与微量滴定板结合的靶蛋白，然后通过ELISA纯化所选噬菌体。另外，还有一种基于珠子的亲和选择方法。这些方法使人们能够轻松分离与感兴趣的蛋白质靶标结合的肽配体，并使用共有序列在蛋白质组数据库中搜索推定的相互作用蛋白。
