NIR-Sensitive Squaraine Dye-Peptide Conjugate for Trypsin Fluorogenic Detection.

IF 4.9 3区 工程技术 Q1 CHEMISTRY, ANALYTICAL
Priyanka Balyan, Shekhar Gupta, Sai Kiran Mavileti, Shyam S Pandey, Tamaki Kato
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Abstract

Trypsin enzyme has gained recognition as a potential biomarker in several tumors, such as colorectal, gastric, and pancreatic cancer, highlighting its importance in disease diagnosis. In response to the demand for rapid, cost-effective, and real-time detection methods, we present an innovative strategy utilizing the design and synthesis of NIR-sensitive dye-peptide conjugate (SQ-3 PC) for the sensitive and selective monitoring of trypsin activity by fluorescence ON/OFF sensing. The current research deals with the design and synthesis of three unsymmetrical squaraine dyes SQ-1, SQ-2, and SQ-3 along with a dye-peptide conjugate SQ-3-PC as a trypsin-specific probe followed by their photophysical characterizations. The absorption spectral investigation conducted on both the dye alone and its corresponding dye-peptide conjugates in water, utilizing SQ-3 and SQ-3 PC respectively, reveals enhanced dye aggregation and pronounced fluorescence quenching compared to observations in DMSO solution. The absorption spectral investigation conducted on dye only and corresponding dye-peptide conjugates in water utilizing SQ-3 and SQ-3 PC, respectively, reveals not only the enhanced dye aggregation but also pronounced fluorescence quenching compared to that observed in the DMSO solution. The trypsin-specific probe SQ-3 PC demonstrated a fluorescence quenching efficiency of 61.8% in water attributed to the combined effect of aggregation-induced quenching (AIQ) and fluorescence resonance energy transfer (FRET). FRET was found to be dominant over AIQ. The trypsin-mediated hydrolysis of SQ-3 PC led to a rapid and efficient recovery of quenched fluorescence (5-fold increase in 30 min). Concentration-dependent changes in the fluorescence at the emission maximum of the dyes reveal that SQ-3 PC works as a trypsin enzyme-specific fluorescence biosensor with linearity up to 30 nM along with the limit of detection and limit of quantification of 1.07 nM and 3.25 nM, respectively.

用于胰蛋白酶荧光检测的近红外敏感水杨酸染料肽共轭物。
胰蛋白酶已被认为是结直肠癌、胃癌和胰腺癌等多种肿瘤的潜在生物标记物,这凸显了它在疾病诊断中的重要性。为了满足对快速、经济、实时检测方法的需求,我们提出了一种创新策略,利用设计和合成近红外敏感染料-肽共轭物(SQ-3 PC),通过荧光开/关传感灵敏、选择性地监测胰蛋白酶活性。目前的研究涉及设计和合成三种非对称方碱染料 SQ-1、SQ-2 和 SQ-3,以及作为胰蛋白酶特异性探针的染料-肽共轭物 SQ-3-PC,并对其进行光物理表征。分别利用 SQ-3 和 SQ-3 PC 对单独的染料及其相应的染料-肽共轭物在水中进行的吸收光谱研究表明,与在二甲基亚砜溶液中观察到的结果相比,染料的聚集性增强,荧光淬灭明显。分别利用 SQ-3 和 SQ-3 PC 对水中的染料和相应的染料-肽共轭物进行的吸收光谱研究表明,与在二甲基亚砜溶液中观察到的结果相比,不仅染料聚集增强,而且荧光淬灭也很明显。由于聚集诱导淬灭(AIQ)和荧光共振能量转移(FRET)的共同作用,胰蛋白酶特异性探针 SQ-3 PC 在水中的荧光淬灭效率达到 61.8%。研究发现,FRET 比 AIQ 起主导作用。胰蛋白酶介导的 SQ-3 PC 水解可使淬灭的荧光快速有效地恢复(30 分钟内增加 5 倍)。染料发射最大值处荧光的浓度变化表明,SQ-3 PC 是一种胰蛋白酶特异性荧光生物传感器,其线性关系可达 30 nM,检测限和定量限分别为 1.07 nM 和 3.25 nM。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Biosensors-Basel
Biosensors-Basel Biochemistry, Genetics and Molecular Biology-Clinical Biochemistry
CiteScore
6.60
自引率
14.80%
发文量
983
审稿时长
11 weeks
期刊介绍: Biosensors (ISSN 2079-6374) provides an advanced forum for studies related to the science and technology of biosensors and biosensing. It publishes original research papers, comprehensive reviews and communications. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced. Electronic files and software regarding the full details of the calculation or experimental procedure, if unable to be published in a normal way, can be deposited as supplementary electronic material.
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