Pillai Sreenadh Sasidharan, Yukawa Hiroshi, Onoshima Daisuke, Biju Vasudevanpillai, Baba Yoshinobu
Graduate School of Engineering, Nagoya University , Furo-cho, Chikusa-Ku, Nagoya , Japan.
† FIRST Research Center for Innovative Nanobiodevices, Nagoya University , Furo-cho, Chikusa-Ku, Nagoya , Japan.
Cell Med. 2015 Aug 26;8(1-2):57-62. doi: 10.3727/215517915X689074. eCollection 2015 Dec 17.
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.
量子点(QDs)最近已被研究用作检测极少量生物分子和活细胞的荧光探针;然而,具有量子点荧光开关控制的分子成像技术仍有待建立。在这里,我们通过使用链霉亲和素-QDs585和生物素-肽-BHQ-1,实现了通过肽介导的黑洞猝灭剂(BHQ)分子与量子点结合,使量子点处于荧光关闭状态。添加生物素-肽-BHQ-1后,链霉亲和素-QDs585的荧光以剂量依赖的方式降低。通过对链霉亲和素-QDs585和QDs585-肽-BHQ-1的荧光强度和寿命分析表明,QDs585荧光的降低是通过Förster共振能量转移(FRET)机制发生的。在该系统中,QDs585荧光的猝灭效率可超过60%。中间肽(pep)的序列为GPLGVRGK,它可被癌细胞产生的基质金属蛋白酶(MMPs)切割。因此,QDs585-肽-BHQ-1有望作为一种用于分子成像的新型生物分析试剂,通过恢复QDs585荧光来检测MMP的产生。