噬菌体展示
计算生物学
肽库
噬菌体
组合化学
肽
淘选
生物
化学
生物化学
基因
肽序列
大肠杆菌
作者
Yu Gu,Jacob A. Iannuzzelli,Rudi Fasan
出处
期刊:Methods in molecular biology
日期:2021-10-02
卷期号:: 261-286
被引量:2
标识
DOI:10.1007/978-1-0716-1689-5_14
摘要
Macrocyclic peptides represent promising scaffolds for targeting biomolecules with high affinity and selectivity, making methods for the diversification and functional selection of these macrocycles highly valuable for drug discovery purposes. We recently reported a novel phage display platform (called MOrPH-PhD) for the creation and functional exploration of combinatorial libraries of genetically encoded cyclic peptides. In this system, spontaneous, posttranslational peptide cyclization by means of a cysteine-reactive non-canonical amino acid is integrated with M13 bacteriophage display, enabling the creation of genetically encoded macrocyclic peptide libraries displayed on phage particles. Using this system, it is possible to rapidly generate and screen large libraries of phage-displayed macrocyclic peptides (up to 108 to 1010 members) in order to identify high-affinity binders of a target protein of interest. Herein, we describe step-by-step protocols for the production of MOrPH-PhD libraries, the screening of these libraries against an immobilized protein target, and the isolation and characterization of functional macrocyclic peptides from these genetically encoded libraries.
科研通智能强力驱动
Strongly Powered by AbleSci AI