利用开放式表面等离子共振(OpenSPR)鉴定蛋白质-蛋白质相互作用的结合亲和力。

Cassie Shu Zhu, Jianhua Li, Haichao Wang
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摘要

表面等离子体共振(SPR)是一种无标记光学技术,用于实时评估蛋白质与蛋白质之间的相互作用动力学和亲和力。传统上,Biacore SPR 采用的是连续的金膜,当连接在平面金表面的配体的折射率因与局部分析物的相互作用而发生变化时,它能检测到再发射光角度的任何变化。相比之下,Nicoya Lifesciences 的 OpenSPR 技术利用金纳米粒子来检测共轭配体与分析物作用后吸光峰波长的微小变化。具体来说,当宽带白光照射到金纳米粒子上时,会产生与金纳米粒子表面共轭配体折射率相对应的强共振吸收峰。然而,在与分析物相互作用时,共轭配体的吸光波长峰会发生变化,并及时记录为配体与分析物相互作用的动态传感图。因此,检测方法的改进(从传统的检测再发射光的角度变化到现代的检测吸光度峰的波长变化)使 OpenSPR 成为深入表征蛋白质-蛋白质相互作用的一种经济高效、操作简便的技术。在这里,我们将详细介绍使用尼科亚生命科学公司的第一代 OpenSPR 仪器(单通道检测)表征原胰蛋白酶 L(pCTS-L)与两种假定的模式识别受体(TLR4 和 RAGE)相互作用的方法。主要特点 - Nicoya OpenSPR 是一种台式小型仪器,可实时深入测量蛋白质-蛋白质相互作用的无标记结合动力学和亲和力。- 该技术相对直观,用户界面友好,适合任何技术水平的科学家使用。- OpenSPR 传感器采用纳米技术,降低了制造复杂光学硬件和传感器芯片的成本,同样也减少了珍贵分析样品的消耗。- 制造商为客户科学家提供 OpenSPR(目录:TRAIN-REMOTE)和 TraceDrawer(目录:TRAIN-TD)的在线培训。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Use of Open Surface Plasmon Resonance (OpenSPR) to Characterize the Binding Affinity of Protein-Protein Interactions.

Surface Plasmon Resonance(SPR) is a label-free optical technique to assess protein-protein interaction kinetics and affinities in a real-time setting. Traditionally, Biacore SPR employs a continuous film of gold to detect any change in the angle of re-emitted light when the refractive index of a ligand conjugated to the flat gold surface is altered by its interaction with a local analyte. In contrast, the Nicoya Lifesciences' OpenSPR technology uses gold nanoparticles to detect small changes in the absorbance peak wavelength of a conjugated ligand after its engagement by an analyte. Specifically, when broadband white light is shone onto the gold nanoparticles, it produces a strong resonance absorbance peak corresponding to the refractive index of a ligand conjugated to the surface of gold nanoparticles. Upon its interaction with an analyte, however, the absorbance wavelength peak of the conjugated ligand will be changed and timely recorded as sensorgrams of dynamic ligand-analyte interactions. Thus, the improvement in the detection method (from traditional detection of changes in the angle of re-emitted light to the contemporary detection of changes in the wavelength of the absorbance peak) features OpenSPR as a cost-effective and user-friendly technique for in-depth characterization of protein-protein interactions. Here, we describe the detailed method that we used to characterize procathepsin L (pCTS-L) interactions with two putative pattern recognition receptors (TLR4 and RAGE) using the 1st generation of Nicoya Lifesciences' OpenSPR instrument with a 1-channel detection. Key features • Nicoya OpenSPR is a benchtop small-size equipment that provides in-depth label-free binding kinetics and affinity measurement for protein-protein interactions in real-time fashion. • This technology is relatively intuitive and user-friendly for scientists at any skill level. • OpenSPR sensors employ nanotechnology to reduce the cost of manufacturing complex optical hardware and Sensor Chips, and similarly reduce the consumption of precious analyte samples. • The manufacturer provides online training for OpenSPR (Catalog: TRAIN-REMOTE) and TraceDrawer (Catalog: TRAIN-TD) to customer scientists.

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