Allen Jessie L, Kennedy Sarah J, Shaw Lindsey N
University of South Florida, Tampa, FL, United States.
University of South Florida, Tampa, FL, United States.
Methods Enzymol. 2022;663:131-156. doi: 10.1016/bs.mie.2021.10.008. Epub 2021 Dec 9.
With the post-antibiotic era rapidly approaching, naturally-sourced antimicrobial peptides (AMPs) are a prime resource for restocking our antibiotic medicine cupboard. The efficiency of identification requires high-throughput screens that can identify bioactive peptides present within abundant natural-products chemical-space. While there are multiple amenable and high sensitivity colorimetric-based screening approaches available, resazurin-based assays are cost-effective, peptide compatible, and expedient, allowing one to screen a multitude of AMPs in a high-throughput fashion. Herein, we provide a detailed protocol for the optimization and use of resazurin assays for AMP testing, providing key experimental insight, and highlight pitfalls to be avoided.
随着后抗生素时代的迅速临近，天然来源的抗菌肽（AMPs）是充实我们抗生素药柜的主要资源。鉴定效率需要高通量筛选，以识别丰富的天然产物化学空间中存在的生物活性肽。虽然有多种适用且高灵敏度的基于比色法的筛选方法，但基于刃天青的测定具有成本效益、与肽兼容且便捷，能够以高通量方式筛选多种抗菌肽。在此，我们提供了用于抗菌肽测试的刃天青测定优化和使用的详细方案，提供关键实验见解，并强调需避免的陷阱。
