Fluorescence Quenching of CdSe/ZnS Quantum Dots by Using Black Hole Quencher Molecules Intermediated With Peptide for Biosensing Application.

Sreenadh Sasidharan Pillai, H. Yukawa, D. Onoshima, V. Biju, Y. Baba
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引用次数: 9

Abstract

Quantum dots (QDs) have recently been investigated as fluorescent probes for detecting a very small number of biomolecules and live cells; however, the establishment of molecular imaging technology with on-off control of QD fluorescence remains to be established. Here we have achieved the fluorescence off state of QDs with the conjugation of black hole quencher (BHQ) molecules intermediated with peptide by using streptavidin-QDs585 and biotin-pep-BHQ-1. The fluorescence of streptavidin-QDs585 was decreased by the addition of biotin-pep-BHQ-1 in a dose-dependent manner. It has been suggested that the decrease in QDs585 fluorescence occurred through a Förster resonance energy transfer (FRET) mechanism from the analysis of fluorescence intensity and lifetime of streptavidin-QDs585 and QDs585-pep-BHQ-1. QDs585 fluorescence could be quenched by more than 60% efficiency in this system. The sequence of intermediate peptide (pep) was GPLGVRGK, which can be cleaved by matrix metalloproteinases (MMPs) produced by cancer cells. QDs585-pep-BHQ-1 is thus expected to detect the MMP production by the recovery of QDs585 fluorescence as a new bioanalytical agent for molecular imaging.
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黑洞猝灭分子介导的CdSe/ZnS量子点荧光猝灭及其生物传感应用
量子点(QDs)最近被研究作为荧光探针来检测极少量的生物分子和活细胞;然而,基于QD荧光开关控制的分子成像技术的建立仍有待建立。本文利用链亲和素- qds585和生物素-pep-BHQ-1,实现了以肽为中间体的黑洞猝灭剂(BHQ)分子偶联的量子点荧光关闭态。生物素-pep- bhq -1的加入使链霉亲和素- qds585的荧光呈剂量依赖性降低。通过对streptavidin-QDs585和QDs585-pep- bhq -1荧光强度和寿命的分析,认为QDs585荧光减弱是通过Förster共振能量转移(FRET)机制发生的。该系统对QDs585荧光的猝灭效率可达60%以上。中间肽(pep)序列为GPLGVRGK,可被癌细胞产生的基质金属蛋白酶(MMPs)裂解。因此,QDs585-pep- bhq -1有望通过QDs585荧光的回收来检测MMP的产生,作为一种新的分子成像生物分析试剂。
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Cell medicine
Cell medicine MEDICINE, RESEARCH & EXPERIMENTAL-
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