State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China.
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, PR China.
Biosens Bioelectron. 2016 Jan 15;75:404-10. doi: 10.1016/j.bios.2015.08.068. Epub 2015 Sep 1.
A benzoperylene probe excimer emission in an aqueous buffer solution is observed for the first time, and a novel ratiometric fluorescence method based on the probe excimer emission for the sensitive detection of heparin and heparinase is demonstrated. A negatively charged benzoperylene derivative, 6-(benzo[ghi]perylene-1,2-dicarboxylic imide-yl)hexanoic acid (BPDI), was employed. A polycation, poly(diallyldimethylammonium) chloride (poly-DDA), could induce aggregation of BPDI through noncovalent interactions. A decrease of BPDI monomer emission and a simultaneous increase of BPDI excimer emission were observed. Upon the addition of heparin, the strong binding between heparin and poly-DDA caused release of BPDI monomer molecules, and an excimer-monomer emission signal transition was detected. However, after the enzymatic hydrolysis of heparin by heparinase, heparin was hydrolyzed into small fragments, which weakened the competitive binding of heparin to poly-DDA. Poly-DDA induced aggregation of BPDI, and a monomer-excimer emission signal transition was detected. Our assay is simple, rapid, inexpensive, sensitive and selective, which could facilitate the heparin and heparinase related biochemical and biomedical research.
首次观察到水溶液中苯并菲探针激基缔合物的发射，并基于探针激基缔合物的发射展示了一种新颖的比率荧光法，用于灵敏检测肝素和肝素酶。使用了带负电荷的苯并菲衍生物 6-(苯并[ghi]菲-1,2-二羧酸酰亚胺基)己酸（BPDI）。聚阳离子聚二烯丙基二甲基氯化铵（poly-DDA）可以通过非共价相互作用诱导 BPDI 的聚集。观察到 BPDI 单体发射的减少和 BPDI 激基缔合物发射的同时增加。当加入肝素时，肝素与 poly-DDA 之间的强结合导致 BPDI 单体分子释放，并检测到激基缔合物-单体发射信号的转变。然而，在肝素酶对肝素的酶解作用下，肝素被水解成小片段，这削弱了肝素与 poly-DDA 的竞争结合。poly-DDA 诱导 BPDI 的聚集，并检测到单体-激基缔合物发射信号的转变。我们的测定方法简单、快速、廉价、灵敏且具有选择性，这将有助于肝素和肝素酶相关的生化和生物医学研究。
