Sultana Azmiri, Lee Jeffrey E
Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, Canada.
Curr Protoc Protein Sci. 2015 Feb 2;79:19.25.1-19.25.26. doi: 10.1002/0471140864.ps1925s79.
Biolayer interferometry (BLI) is a simple, optical dip-and-read system useful for measuring interactions between proteins, peptides, nucleic acids, small molecules, and/or lipids in real time. In BLI, a biomolecular bait is immobilized on a matrix at the tip of a fiber-optic sensor. The binding between the immobilized ligand and another molecule in an analyte solution produces a change in optical thickness at the tip and results in a wavelength shift proportional to binding. BLI provides direct binding affinities and rates of association and dissociation. This unit describes an efficient approach using streptavidin-based BLI to analyze DNA-protein and protein-protein interactions. A quantitative set of equilibrium binding affinities (K(d)) and rates of association and dissociation (k(a)/k(d)) can be measured in minutes using nanomole quantities of sample.
生物层干涉术（BLI）是一种简单的光学浸入式读数系统，可用于实时测量蛋白质、肽、核酸、小分子和/或脂质之间的相互作用。在BLI中，生物分子诱饵固定在光纤传感器尖端的基质上。固定化配体与分析物溶液中的另一种分子之间的结合会导致尖端光学厚度发生变化，并产生与结合成正比的波长偏移。BLI可提供直接的结合亲和力以及缔合和解离速率。本单元介绍了一种使用基于链霉亲和素的BLI分析DNA-蛋白质和蛋白质-蛋白质相互作用的有效方法。使用纳摩尔量的样品，可在数分钟内测量出一组定量的平衡结合亲和力（K(d)）以及缔合和解离速率（k(a)/k(d)）。
