基于小分子末端保护的turn-on传感器构建及其在蛋白质检测中的应用  

″Turn-on″ Fluorescent Sensor for Protein Determination Based on Terminal Protection of Small-Molecule-Linked DNA

作　　者：陈朝会 张正涛 刘小云 CHEN Chaohui;ZHANG Zhengtao;LIU Xiaoyun(Interdisciplinary Research Institute,Jianghan University,Wuhan 430056,Hubei,China)

机构地区：[1]江汉大学交叉学科研究院

出　　处：《江汉大学学报（自然科学版）》2019年第5期442-448,共7页Journal of Jianghan University：Natural Science Edition

基　　金：湖北省教育厅科学技术研究计划指导性项目(B2017264)

摘　　要：基于小分子末端保护技术,结合原位合成DNA功能化银簇,构建了一种简单且低成本的蛋白质检测方法。本方法检测链霉亲和素的线性范围为10~1000 ng/mL,检出限为7.92 ng/mL,具有较好的选择性并且可以实现复杂样品中目标蛋白质的检测;此外,通过在探针核酸一端修饰上不同的小分子可用于其他相对应蛋白质和细胞的检测,具有良好的通用性;核酸外切酶I的引入,将没有与目标蛋白结合的探针DNA水解为单核苷酸,使之不能形成DNA功能化的银簇,从而降低背景干扰,提高灵敏度;该探针无需修饰荧光染料,且合成银簇所需原料廉价易得,大大节约了成本。A simple and low-cost method for the determination of protein was established.This method was based on the terminal protection of small-molecule-linked DNA and used the DNAstabilized Ag nanoclusters(DNA-Ag NC).This sensor could detect the target protein streptavidin(SA)as low as 7.92 ng/mL with a linear range from 10 to 1 000 ng/mL and exhibited good selectivity.Besides,this method could be used as a universal method for protein and cell determination by changing the corresponding small molecule.More importantly,the introduction of Exonuclease I(Exo I)hydrolyze probe DNA that didn′t bind to a target protein into mononucleotides,thus the DNA-Ag NC could not be formed,the background interference was reduced and the sensitivity was improved.Furthermore,the probe did not need to modify fluorescent dyes,and the raw materials for synthesizing silver clusters were cheap and easy to obtain,which greatly saved the cost.

关 键 词：银簇 小分子末端保护 酶解作用 生物传感 链霉亲和素 

分 类 号：O652[理学—分析化学]