A cationic conjugated polymer coupled with exonuclease I: application to the fluorometric determination of protein and cell imaging

文献类型： 外文期刊

第一作者： Liu, Yufei

作者： Liu, Yufei;Yan, Huijuan;Shangguan, Jingfang;Gao, Liyun;Zhang, Zhen;Xiang, Xia

作者机构：

关键词： PFP;FRET;Streptavidin;Folate receptor;TPSMLD;Immunoglobulin G

期刊名称：MICROCHIMICA ACTA （ 影响因子：5.833； 五年影响因子：5.357 ）

ISSN： 0026-3672

年卷期： 2018 年 185 卷 2 期

页码：

收录情况： SCI

摘要： A strategy is described for the detection of protein by using a cationic fluorescent conjugated polymer coupled with exonuclease I (Exo I). Taking streptavidin (SA) as model protein, it is observed that Exo I can digest single-stranded DNA conjugated with biotin and carboxyfluorescein (P1) if SA is absent. This leads to the formation of small nucleotide fragments and to weak fluorescence resonance energy transfer (FRET) from the polymer to P1. If, however, SA is present, the high affinity of SA and biotin prevents the digestion of P1 by Exo I. This results in the sorption of P1 on the surface of the polymer through strong electrostatic interaction. Hence, efficient FRET occurs from the fluorescent polymer to the fluorescent label of P1. Fluorescence is measured at an excitation wavelength of 370 nm, and emission is measured at two wavelengths (530 and 425 nm). The ratio of the two intensities (I-530/I-425) is directly related to the concentration of SA. Under the optimal conditions, the assay has a detection limit of 1.3 ng.mL(-1). The method was also applied to image the folate receptor in HeLa cells, thus demonstrating the versatility of this strategy.

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