Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi'an 710062, China.
Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi'an 710062, China.
Talanta. 2017 Dec 1;175:224-228. doi: 10.1016/j.talanta.2017.07.003. Epub 2017 Jul 6.
A colorimetric assay for the ultrasensitive determination of thrombin was presented, in which the cationic polythiophene derivative was used as catalyst of the 3,3',5,5'-tetramethylbenzidine (TMB)-HO reaction and the thrombin-binding aptamer (TBA) was used as inducing polymer's different conformation elements. It was found the cationic polythiophene derivative, poly[3-(3'-N,N,N-triethylamino-1'-propyloxy)-4-methyl-2,5-t-hiophene hydrochloride] (PMNT), can catalyze the oxidation reaction of TMB in the presence of HO to produce a blue color solution. The catalytic activity of PMNT on the TMB-HO reaction was closely relevant to the conformation of PMNT. The absorbance of TMB-HO was distinctly increased in the presence of TBA. With the addition of thrombin, TBA interacted with thrombin to form a G-quadruplex structure. The conformational change weakened the catalytic activity of PMNT and resulted in a decrease in the absorbance. The colorimetric sensor could detect thrombin down to 4pM with high selectivity against other interfering proteins. This work is not only of importance for a better understanding of the unique properties of cationic polythiophenes derivative but also have great potential for medical diagnostics and therapy for human health.
本文提出了一种用于超灵敏检测凝血酶的比色分析方法,其中阳离子聚噻吩衍生物被用作 3,3',5,5'-四甲基联苯胺 (TMB)-HO 反应的催化剂,凝血酶结合适体 (TBA) 被用作诱导聚合物不同构象的元素。研究发现,阳离子聚噻吩衍生物聚[3-(3'-N,N,N-三乙基氨基-1'-丙氧基)-4-甲基-2,5-噻吩盐酸盐] (PMNT) 在 HO 存在下可以催化 TMB 的氧化反应,生成蓝色溶液。PMNT 对 TMB-HO 反应的催化活性与 PMNT 的构象密切相关。在 TBA 的存在下,TMB-HO 的吸光度明显增加。随着凝血酶的加入,TBA 与凝血酶相互作用形成 G-四链体结构。构象的变化削弱了 PMNT 的催化活性,导致吸光度降低。比色传感器可以检测到低至 4pM 的凝血酶,对其他干扰蛋白具有高选择性。这项工作不仅对更好地理解阳离子聚噻吩衍生物的独特性质具有重要意义,而且对医学诊断和人类健康的治疗也具有巨大的潜力。