Young Tracy A, Moree Ben, Butko Margaret T, Clancy Bason, Geck Do Mary, Gheyi Tarun, Strelow John, Carrillo Juan J, Salafsky Joshua
Biodesy, Inc., South San Francisco, CA, United States.
The University of Texas MD Anderson Cancer Center, Institute for Applied Cancer Science, Houston, TX, United States.
Methods Enzymol. 2018;610:167-190. doi: 10.1016/bs.mie.2018.09.017. Epub 2018 Oct 19.
Second-harmonic generation (SHG) has recently emerged as a biophysical tool for conformational sensing of a target biomolecule upon binding to ligands such as small molecules, fragments, proteins, peptides, and oligonucleotides. To date, SHG has been used to measure conformational changes of targets such as soluble proteins, protein complexes, intrinsically disordered proteins, peripheral and integral membrane proteins, peptides, and oligonucleotides upon binding of ligands over a wide range of affinities. In this chapter, we will provide a technology overview, detailed protocols for optimizing assays and screening, practical considerations, and an example case study to guide the reader in developing robust and informative measurements using the Biodesy Delta SHG platform.
二次谐波产生（SHG）最近已成为一种生物物理工具，用于在目标生物分子与小分子、片段、蛋白质、肽和寡核苷酸等配体结合时进行构象传感。迄今为止，SHG已被用于测量多种亲和力范围内配体结合后，可溶性蛋白质、蛋白质复合物、内在无序蛋白质、外周和整合膜蛋白、肽和寡核苷酸等目标的构象变化。在本章中，我们将提供技术概述、优化检测和筛选的详细方案、实际注意事项以及一个案例研究，以指导读者使用Biodesy Delta SHG平台进行可靠且信息丰富的测量。
