Clow Fiona, Fraser John D, Proft Thomas
Department of Molecular Medicine & Pathology, School of Medical Sciences, University of Auckland, Private Bag 92019, Auckland, New Zealand.
Biotechnol Lett. 2008 Sep;30(9):1603-7. doi: 10.1007/s10529-008-9718-1. Epub 2008 Apr 15.
The immobilization of proteins to surfaces is an active area of research due to strong interest in protein-based sensors. Here, we describe a novel method for immobilizing ligand proteins onto Biacore sensor chips using the transpeptidase activity of Staphylococcus aureus sortase A (SrtA). This method provides a robust and gentle approach for the site-directed, covalent coupling of proteins to biosensor chips. Notably, the high specificity of the sortase allows immobilization of proteins from less than pure protein samples allowing short cuts in protein purification protocols.
由于对基于蛋白质的传感器有着浓厚兴趣,蛋白质在表面的固定化是一个活跃的研究领域。在此,我们描述了一种利用金黄色葡萄球菌分选酶A(SrtA)的转肽酶活性将配体蛋白固定到Biacore传感器芯片上的新方法。该方法为蛋白质与生物传感器芯片的定点共价偶联提供了一种稳健且温和的途径。值得注意的是,分选酶的高特异性使得能够从纯度不高的蛋白质样品中固定蛋白质,从而简化蛋白质纯化方案。