基于多肽微阵列芯片的荧光和共振光散射法筛选凝血酶抑制剂

发布时间：2018-04-24 13:43

  本文选题：多肽微阵列 + 金纳米粒子探针　； 参考：《分析化学》2015年02期

【摘要】：研制了一种基于多肽微阵列芯片的荧光和共振光散射双通路检测方法,对血液样品中的凝血酶抑制剂进行检测。以生物素化的多肽微阵列芯片为反应平台,加入凝血酶水解多肽上的特异位点使其C末端的生物素解离,通过亲和素和生物素之间的特异性结合,用荧光探针和30 nm金纳米粒子探针标记此反应过程。凝血酶抑制剂阻止凝血酶对底物多肽的水解反应,通过荧光和共振光散射信号强度的变化检测抑制剂的抑制能力。酶溶液和加标人血清中测定的半抑制浓度(IC50)值:阿加曲班抗凝血酶Ⅲ4-(2-氨乙基)苯磺酰氟盐酸盐,IC50差的绝对值随抑制剂特异性的减弱而增大。当抑制剂浓度为7.5μmol/L时,血浆中5种化合物的抑制能力:阿加曲班抗凝血酶Ⅲ胰蛋白酶抑制剂N-(反式-环氧丁二酰基)-L-亮氨酸-4-胍基丁基酰胺4-(2-氨乙基)苯磺酰氟盐酸盐。在血浆中考察了阿加曲班和抗凝血酶Ⅲ的可逆性。对比荧光法和共振光散射法在血液样品中的检测结果,用30 nm金纳米粒子标记的共振光散射法更适用于复杂血液样品中抑制剂的检测
[Abstract]:A dual fluorescence and resonance light scattering (RLS) method based on polypeptide microarray was developed to detect thrombin inhibitors in blood samples. Using biotin microarray microarray as the reaction platform, the specific site on the thrombin hydrolyzed polypeptide was added to dissociate the biotin from the C-terminal of the peptide, and the specific binding between avidin and biotin was obtained. Fluorescence probe and 30 nm gold nanoparticles probe were used to label the reaction process. Thrombin inhibitor blocked the hydrolysis of the substrate polypeptide by thrombin and detected the inhibition ability of the inhibitor by the changes of fluorescence and resonance light scattering (RLS) signal intensity. The absolute value of IC50 difference of agatoban antithrombin 鈪，

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