Selection and Identification of a DNA Aptamer That Mimics Saxitoxin in Antibody Binding

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• 作者：Pan Hu ; Zengshan Liu ; Ruiyun Tian ; Honglin Ren ; Xiaoxu Wang ; Chao Lin ; Sheng Gong ; Xianmei Meng ; Guangming Wang ; Yu Zhou ; Shiying Lu
• 刊名：Journal of Agricultural and Food Chemistry
• 出版年：2013
• 出版时间：April 10, 2013
• 年：2013
• 卷：61
• 期：14
• 页码：3533-3541
• 全文大小：495K
• 年卷期：v.61,no.14(April 10, 2013)
• ISSN：1520-5118

文摘

In this article, high-affinity single-stranded DNA (ssDNA) aptamer-targeting F(ab鈥?₂ fragments of saxitoxin (STX) antibodies were selected from a random ssDNA library by the SELEX strategy. After 16 rounds of repeated selection, the enriched ssDNA library was sequenced, and all of the sequences were carefully identified by indirect enzyme-linked assay and indirect competitive enzyme-linked assay (icELISA). The candidate aptamers in the above identification were selected for further characterization by icELISA and the equilibrium filtration method. We successfully obtained an aptamer that mimics STX in antibody binding, and a substitute for STX in aptamer form has been developed. Further work is in progress aimed at using this aptamer substitute to replace the STX standard in an antibody-based, nontoxic detection method for field determination of STX in seafood products.

Keywords:

saxitoxin; aptamer; F(ab鈥?₂ fragments; SELEX; paralytic shellfish poisoning