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通过鱼精蛋白诱导的碳量子点猝灭聚集平台对胰蛋白酶进行的简单荧光测定法。

A simple fluorescence assay for trypsin through a protamine-induced carbon quantum dot-quenching aggregation platform.

作者信息

Chen Yiping, Lin Zuan, Miao Chenfang, Cai Qianqian, Li Fenglan, Zheng Zongfu, Lin Xinhua, Zheng Yanjie, Weng Shaohuang

机构信息

Department of Interventional Radiology, The First Affiliated Hospital of Fujian Medical University Fuzhou 350005 China.

Department of Pharmaceutical Analysis, School of Pharmacy, The Higher Educational Key Laboratory for Nano Biomedical Technology of Fujian Province, Fujian Medical University Fuzhou 350122 China

出版信息

RSC Adv. 2020 Jul 28;10(45):26765-26770. doi: 10.1039/d0ra03970e. eCollection 2020 Jul 15.

Abstract

The development of a simple detection strategy for trypsin (Try) is urgent, and is ascribed to the diagnostic value of Try in several diseases. Herein, a facile but effective fluorescence strategy for Try was developed based on the protamine (Pro)-induced aggregation of carbon quantum dots (CQDs). The fluorescence of negatively charged CQDs was quenched with Pro due to the assembly of CQDs and Pro (CQDs/Pro) through electrostatic interaction. However, the highly positively charged Pro, which is rich in basic arginine residues, was preferred to be hydrolyzed by Try. Try can induce the deaggregation of CQDs/Pro, thereby enabling the release of CQDs to restore the fluorescence intensity. Thus, the use of CQDs/Pro as a testing platform will be employed as a "turn-on" method for Try. In addition, the fluorescence-resuming response was proportional to Try, ranging from 25 ng mL to 500 ng mL with a limit of detection (LOD) of 8.08 ng mL. This "turn-on" fluorescence assay for Try was label-free, convenient, and relatively free of interference from coexisting substances. Actual applications for Try monitoring and trypsin inhibitor screening also illustrated the considerable prospect of CQDs in the clinical field, combined with the superiority of the simple mixing operation.

摘要

开发一种简单的胰蛋白酶(Try)检测策略迫在眉睫,这归因于Try在多种疾病中的诊断价值。在此,基于鱼精蛋白(Pro)诱导的碳量子点(CQDs)聚集,开发了一种简便而有效的检测Try的荧光策略。带负电荷的CQDs的荧光由于CQDs与Pro通过静电相互作用组装形成CQDs/Pro而被淬灭。然而,富含碱性精氨酸残基的带高度正电荷的Pro更易被Try水解。Try可诱导CQDs/Pro解聚,从而使CQDs释放出来恢复荧光强度。因此,将CQDs/Pro用作检测平台将作为一种检测Try的“开启”方法。此外,荧光恢复响应与Try成正比,范围为25 ng/mL至500 ng/mL,检测限(LOD)为8.08 ng/mL。这种检测Try的“开启”荧光分析法无需标记、操作简便,且相对不受共存物质的干扰。对Try监测和胰蛋白酶抑制剂筛选的实际应用也说明了CQDs结合简单混合操作的优越性在临床领域具有广阔前景。

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