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基于金纳米粒子增强的辣根过氧化物酶信号放大策略的降钙素原超灵敏电化学免疫传感器。

An ultrasensitive electrochemical immunosensor for procalcitonin detection based on the gold nanoparticles-enhanced tyramide signal amplification strategy.

机构信息

State Key Laboratory of Pharmaceutical Biotechnology and The Comprehensive Cancer Center, Nanjing Drum Tower Hospital, MOE Key Laboratory of Model Animal for Disease Study, School of Life Sciences, Nanjing University, Nanjing 210046, PR China.

State Key Laboratory of Pharmaceutical Biotechnology and Collaborative Innovation Center of Chemistry for Life Sciences, School of Life Sciences, Nanjing University, Nanjing 210046, PR China.

出版信息

Biosens Bioelectron. 2019 Feb 1;126:543-550. doi: 10.1016/j.bios.2018.10.048. Epub 2018 Oct 28.

Abstract

In this study, we established an ultrasensitive electrochemical immunosensor based on the gold nanoparticles-enhanced tyramide signal amplification (AuNPs-TSA) for the detection of procalcitonin (PCT, for discriminating bacterial infections from nonbacterial infections). Firstly, a facilely prepared, well-conducting reduced graphene oxide nanosheets/GNP (rGO-AuNPs) nanocomposite was synthesized and immobilized on the electrode surface to absorb more capture antibodies (Ab1). Next another nanocomposite, acting as a signal tool, was modified with detection antibody (Ab2) and horseradish peroxidase (HRP), and then backfilled by bovine serum albumin (BSA). Because a single AuNP is able to load multiple HRPs and BSAs, a number of tyramine labeled biotins (T-B) could be deposited on the proteins adhering to the surface of AuNPs. Moreover, the high affinity between streptavidin (SA) and biotins significantly increases the loading of streptavidin labeled horseradish peroxidase (SA-HRP). The amplification system which was based on the two nanocomposites mentioned above, effectively amplified the electric current signals. This immunosensor exhibits a wide dynamic detection range from 0.05 ng mL to 100 ng mL and with an ultralow detection limit of 0.1 pg mL. We have successfully utilized this immunosensor to quantify the concentration of PCT in human serum samples, and the results suggest its potential use in clinical application.

摘要

在本研究中,我们建立了一种基于金纳米粒子增强的辣根过氧化物酶信号放大(AuNPs-TSA)的超灵敏电化学免疫传感器,用于检测降钙素原(PCT,用于区分细菌性感染和非细菌性感染)。首先,我们制备了一种简便的、导电性良好的还原氧化石墨烯纳米片/纳米金(rGO-AuNPs)纳米复合材料,并将其固定在电极表面以吸附更多的捕获抗体(Ab1)。接下来,另一种纳米复合材料被修饰上检测抗体(Ab2)和辣根过氧化物酶(HRP),并进一步用牛血清白蛋白(BSA)进行封闭。由于单个金纳米粒子能够负载多个 HRP 和 BSA,因此可以将大量的标记有辣根过氧化物酶的生物素(T-B)沉积在附着于 AuNPs 表面的蛋白质上。此外,链霉亲和素(SA)和生物素之间的高亲和力显著增加了 SA 标记的辣根过氧化物酶(SA-HRP)的负载量。基于上述两种纳米复合材料的放大系统有效地放大了电流信号。该免疫传感器具有从 0.05ng/mL 到 100ng/mL 的宽动态检测范围,检测限低至 0.1pg/mL。我们成功地利用该免疫传感器对人血清样品中的 PCT 浓度进行了定量检测,结果表明其在临床应用中的潜在用途。

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