Geological Research Institute for Coal Green Mining, College of Geology and Environment, Xi'an University of Science and Technology, Xi'an 710054, PR China.
Geological Research Institute for Coal Green Mining, College of Geology and Environment, Xi'an University of Science and Technology, Xi'an 710054, PR China.
Spectrochim Acta A Mol Biomol Spectrosc. 2019 Jun 5;216:310-318. doi: 10.1016/j.saa.2019.03.073. Epub 2019 Mar 19.
Heparin is a glycosaminoglycan with the highest negative charge density of any known biological molecule. Herein, this highly negative charge structure of heparin and the charge effect from positively-charged AuNPs for luminol chemiluminescence (CL) reaction were combined to build a facile and sensitive CL strategy for detection of heparin. The highly negative charge structure of heparin molecules (four negatively-charged side groups per repeat unit) and the effective signal amplification of charge effect from positively-charged AuNPs make this analysis to display high sensitivity for heparin detection, and the detection limit is as low as 0.06 ng/mL. It is about two orders of magnitude lower than the previously reported colorimetric assay and far lower than the current analysis methods. The established CL strategy is to use the electrostatic interaction between heparin and signal probe (positively-charged AuNPs). Since polyanionic heparin has the highest negative charge in biological system, this CL sensing shows high selectivity for the detection of heparin, and hyaluronic acid (HA), an analogue of heparin, cannot cause interference. This CL sensing succeeded in detecting heparin in human serum samples. Besides, polycationic protamine, heparin antidote, can respond to the system's CL signals through its strong interactions with heparin, thus indirectly detecting protamine. For protamine in serum samples, the detection result was basically consistent with Coomassie brilliant blue assay.
肝素是一种糖胺聚糖，其负电荷密度是任何已知生物分子中最高的。在此，将肝素的这种高负电荷结构与带正电荷的 AuNPs 的电荷效应结合起来，用于构建鲁米诺化学发光（CL）反应的简便、灵敏的 CL 策略，以检测肝素。肝素分子的高负电荷结构（每个重复单元有四个带负电荷的侧基）和带正电荷的 AuNPs 的电荷效应的有效信号放大，使得该分析对肝素检测具有高灵敏度，检测限低至 0.06ng/mL。这大约比以前报道的比色测定低两个数量级，远低于目前的分析方法。所建立的 CL 策略是利用肝素与信号探针（带正电荷的 AuNPs）之间的静电相互作用。由于肝素在生物体系中带有的负电荷最高，因此这种 CL 感测对肝素的检测具有高选择性，而肝素类似物透明质酸（HA）不能引起干扰。该 CL 感测成功地检测了人血清样品中的肝素。此外，肝素解毒剂聚阳离子鱼精蛋白可以通过与肝素的强相互作用来响应系统的 CL 信号，从而间接检测鱼精蛋白。对于血清样品中的鱼精蛋白，检测结果与考马斯亮蓝测定基本一致。
