Magnetic Separation-Based Multiple SELEX for Effectively Selecting Aptamers against Saxitoxin, Domoic Acid, and Tetrodotoxin

Journal of Agricultural and Food Chemistry
Huajie GuZhouping Wang

Abstract

In this study, a novel magnetic separation-based multiple systematic evolution of ligands by exponential enrichment (SELEX) was applied to select aptamers simultaneously against three kinds of marine biotoxins, including domoic acid (DA), saxitoxin (STX), and tetrodotoxin (TTX). Magnetic reduced graphene oxide (MRGO) was prepared to adsorb unbound ssDNAs and simplify the separation step. In the multiple SELEX, after the initial twelve rounds of selection against mixed targets and the subsequent four respective rounds of selection against each single target, the three resulting ssDNA pools were cloned, sequenced, and analyzed. Several aptamer candidates were selected and subjected to the binding affinity and specificity test. Finally, DA-06 ( Kd = 62.07 ± 19.97 nM), TTX-07 ( Kd = 44.12 ± 15.38 nM), and STX-41 ( Kd = 61.44 ± 23.18 nM) showed high affinity and good specificity for DA, TTX, and STX, respectively. They were also applied to detect and quantify DA, TTX, and STX successfully. The other two multitarget aptamers, DA-01 and TTX-27, were also obtained, which can bind with either DA or TTX. These aptamers provide alternative recognition molecules to antibodies for biosensor applications.

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Citations

Aug 14, 2020·PloS One·Boutheina MarnissiIssam Hmila
May 6, 2020·International Journal of Molecular Sciences·Tin Hoang Trung ChauEun Yeol Lee
Nov 3, 2020·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Shingo Saito
Jun 19, 2021·Food Chemistry·Monali MukherjeePraveena Bhatt
Dec 7, 2019·Journal of Agricultural and Food Chemistry·Jee Young KimYoon-E Choi
Aug 25, 2021·Biotechnology and Applied Biochemistry·Michael Kohlberger, Gabriele Gadermaier
Oct 27, 2021·Analytical Chemistry·Xhensila ShkembiCiara K O Sullivan
Dec 3, 2021·Critical Reviews in Food Science and Nutrition·Mingqi ZhaoA M Abd El-Aty

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