Simple and Efficient Method to Purify DNA鈥揚rotein Conjugates and Its Sensing Applications

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• 作者：Zhaojuan Zhou ; Yu Xiang ; Aijun Tong ; Yi Lu
• 刊名：Analytical Chemistry
• 出版年：2014
• 出版时间：April 15, 2014
• 年：2014
• 卷：86
• 期：8
• 页码：3869-3875
• 全文大小：374K
• 年卷期：v.86,no.8(April 15, 2014)
• ISSN：1520-6882

文摘

DNA鈥損rotein conjugates are very useful in analytical chemistry for target recognition and signal amplification. While a number of methods for conjugating DNA with proteins are known, methods for purification of DNA鈥損rotein conjugates from reaction mixture containing unreacted proteins are much less investigated. In this work, a simple and efficient approach to purify DNA鈥搃nvertase conjugates from reaction mixture via a biotin displacement strategy to release desthiobiotinylated DNA鈥搃nvertase conjugates from streptavidin-coated magnetic beads was developed. The conjugates purified by this approach were utilized for quantitative detection of cocaine and DNA using a personal glucose meter through structure-switching DNA aptamer sensors and competitive DNA hybridization assays, respectively. In both cases, the purified DNA鈥搃nvertase conjugates showed better performance compared to the same assays using unpurified conjugates. The approach demonstrated here can be further expanded to other DNA and proteins to generate purified DNA鈥損rotein conjugates for analytical and other applications.