Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Naval Medical University, Shanghai 200433, China.
Toxins (Basel). 2022 Oct 14;14(10):706. doi: 10.3390/toxins14100706.
Conotoxins (CTXs) are a variety of mixed polypeptide toxins, among which α-conotoxin MI (CTX-MI) is the most toxic. Serious toxic symptoms, a lack of counteracting drugs, and cumbersome detection processes have made CTX-MI a hidden danger for humans. One of the obstacles to resolving this problem is the absence of specific recognition elements. Aptamers have shown great advantages in the fields of molecule detection, drug development, etc. In this study, we screened and characterized aptamers for CTX-MI through a programmed process. MBMI-01c, the isolated aptamer, showed great affinity, with an affinity constant (K) of 0.524 μM, and it formed an antiparallel G-quadruplet (GQ) structure for the specific recognition of CTX-MI. Additionally, an aptasensor based on the biolayer interferometry (BLI) platform was developed and displayed high precision, specificity, and repeatability with a limit of detection (LOD) of 0.26 μM. This aptasensor provides a potential tool for the rapid detection of CTX-MI in 10 min. The aptamer can be further developed for the enrichment, detoxification, and biological studies of CTX-MI. Additionally, the programmed process is applicable to screening and characterizing aptamers for other CTXs.
短肽毒素(CTXs)是一类混合多肽毒素,其中α-短肽毒素 MI(CTX-MI)的毒性最强。严重的中毒症状、缺乏解毒药物和繁琐的检测过程使 CTX-MI 成为人类的潜在威胁。解决这一问题的障碍之一是缺乏特异性识别元件。适体在分子检测、药物开发等领域显示出巨大的优势。在本研究中,我们通过程序化过程筛选和鉴定了针对 CTX-MI 的适体。分离出的适体 MBMI-01c 具有很强的亲和力,亲和常数(K)为 0.524 μM,并且它形成了一个反平行 G-四联体(GQ)结构,用于对 CTX-MI 的特异性识别。此外,还基于生物层干涉(BLI)平台开发了一种适体传感器,该传感器具有高精确度、特异性和重复性,检测限(LOD)为 0.26 μM。该适体传感器提供了一种在 10 分钟内快速检测 CTX-MI 的潜在工具。该适体可以进一步开发用于 CTX-MI 的富集、解毒和生物学研究。此外,程序化过程适用于筛选和鉴定其他 CTX 类的适体。