适体
化学
指数富集配体系统进化
生物信息学
离解常数
互补性(分子生物学)
互补决定区
组合化学
纳米技术
计算生物学
核糖核酸
分子生物学
生物化学
受体
生物
基因
肽序列
遗传学
材料科学
作者
Qionglin Wang,Pengbo Guo,Weyland Cheng,Yuchun Liu,Yaodong Zhang,Peng Huo,Shubin Feng,Wancun Zhang
出处
期刊:Talanta
[Elsevier]
日期:2024-03-01
卷期号:269: 125535-125535
标识
DOI:10.1016/j.talanta.2023.125535
摘要
Numerous aptamers against various targets have been identified through the technology of systematic evolution of ligands by exponential enrichment (SELEX), but the affinity of these aptamers are often insufficient due to the limitations of SELEX. Therefore, a more rational in silico screening strategy (ISS) was developed for efficient screening of high affinity aptamers, which took shape complementarity and thermodynamic stability into consideration. Neuron specific enolase (NSE), a tumor marker, was selected as the target molecule. In the screening process, three aptamer candidates with good shape complementarity, lower ΔG values, and higher ZDOCK scores were produced. The dissociation constant (Kd) of these candidates to NSE was determined to be 10.13 nM, 14.82 nM, and 2.76 nM, respectively. Each of them exhibited higher affinity to NSE than the parent aptamer (Kd = 23.83 nM). Finally, an antibody-free fluorescence aptasensor assay, based on the aptamer with the highest affinity, P–5C8G, was conducted, resulting in a limit of detection (LOD) value of 1.8 nM, which was much lower than the parental aptamer (P, LOD = 12.6 nM). The proposed ISS approach provided an efficient and universal strategy to improve the aptamer to have a high affinity and good analytical utility.
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