The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia.
Monash Institute of Pharmaceutical Sciences, Monash University, Melbourne, VIC, Australia.
Methods Mol Biol. 2023;2705:199-210. doi: 10.1007/978-1-0716-3393-9_10.
Biosensor measurement using surface plasmon resonance enables precise evaluation of peptide-protein interactions. It is a sensitive technique that provides kinetic and affinity data with very little sample and without the need for analyte labels. Here, we describe its application for the analysis of peptide interactions with the Grb7-SH2 domain prepared with a GST-tag for tethering to the chip surface. This has been successfully and reliably used for direct comparison of a range of peptides under different solution conditions as well as direct comparison of peptides flowed over different related SH2 domains in real time. We have used the BIAcore system and describe both the methodology for data collection and analysis, with principles also applicable to other biosensor platforms.
生物传感器测量使用表面等离子体共振能够精确评估肽-蛋白质相互作用。这是一种灵敏的技术，提供动力学和亲和力数据，只需很少的样本，而不需要分析物标记。在这里，我们描述了它在分析与 Grb7-SH2 结构域相互作用的肽的应用，该结构域带有 GST 标签用于固定在芯片表面上。这种方法已成功可靠地用于在不同溶液条件下直接比较一系列肽，以及实时直接比较流过不同相关 SH2 结构域的肽。我们使用了 BIAcore 系统，并描述了数据收集和分析的方法，其原理也适用于其他生物传感器平台。
