适体是通过SELEX(通过指数富集的配体的系统进化)选择的单链寡核苷酸,能够以高亲和力和特异性区分靶分子,即使在非常密切相关的结构的情况下。已经为几种靶标产生了适体,包括小分子,如霉菌毒素;然而,对它们各自的靶分子的高亲和力是关键的要求。在过去的十年里,通过计算方法筛选适体对特定靶标的亲和力已大大增加,并且由于其便利性和低成本而成为常用的程序。本文介绍了一种用于快速筛选针对伏马菌素B1(FB1,n=3)的十个ssDNA适体序列的计算机方法,黄曲霉毒素B1(AFB1,n=2)和曲霉毒素A(OTA,n=5)。将理论结果与通过荧光微尺度热泳法和磁珠测定其结合亲和力(KD)测试相同适体所获得的结果进行比较,显示出良好的一致性。
Aptamers are single-stranded oligonucleotides selected by SELEX (Systematic Evolution of Ligands by EXponential Enrichment) able to discriminate target molecules with high affinity and specificity, even in the
case of very closely related structures. Aptamers have been produced for several targets including small molecules like mycotoxins; however, the high affinity for their respective target molecules is a critical requirement. In the last decade, the screening through computational methods of aptamers for their affinity against specific targets has greatly increased and is becoming a commonly used procedure due to its convenience and low costs. This paper describes an in-silico approach for rapid screening of ten ssDNA aptamer sequences against fumonisin B1 (FB1, n = 3), aflatoxin B1 (AFB1, n = 2) and ochratoxin A (OTA, n = 5). Theoretical results were compared with those obtained by testing the same aptamers by fluorescent microscale thermophoresis and by magnetic beads assay for their binding affinity (KD) revealing a good agreement.