核酸适配体结合亲和力评测方法的研究进展  

Research progress on evaluation methods of binding affinity of nucleic acid aptamer

作　　者：汪凯 李延生 高国伟[1,2] 宋钰 胡敬芳 Kai Wang;Yansheng Li;Guowei Gao;Yu Song;Jingfang Hu(Key Laboratory of Sensors,Beijing Information Science and Technology University,Beijing 100101,China;Key Laboratory of Modern Measurement and Control Technology of Ministry of Education,Beijing Information Science and Technology University,Beijing 100192,China)

机构地区：[1]北京信息科技大学传感器重点实验室,北京100101 [2]北京信息科技大学现代测控技术教育部重点实验室,北京100192

出　　处：《中国科学:化学》2025年第2期444-462,共19页SCIENTIA SINICA Chimica

基　　金：国家自然科学基金(62101053);北京市教育委员会科学研究计划(KM202211232004);中央级公益性科研院所基本科研业务费专项资金(RXRC2022001);北京市属高等学校高水平科研创新团队建设支持计划(BPHR20220124)资助项目。

摘　　要：可体外合成的核酸适配体在生命健康领域受到越来越多的关注,其与靶标的结合亲和力是重要的性能指标之一.评测核酸适配体结合亲和力的方法多种多样,我们对技术方法进行筛选分类,详细阐述了每种技术方法的基本原理、研究进展以及需要突破的瓶颈问题.成熟的核酸适配体结合亲和力评测方法涵盖了表面等离子体共振法、生物层干涉法、酶联适体测定法以及石英晶体微天平法等.这些技术能够稳定且灵敏地捕捉分子结合信号,从而显著推动核酸适配体技术的研究进展.然而,传统的核酸适配体固定技术可能会对分子构象产生一定影响,从而改变其在使用环境中的真实结合亲和力.为解决这一问题,研究者们正积极探索新型的无需固定就能评估分子结合亲和力的方法,例如圆二色性法、紫外-可见吸收光谱法、荧光各向异性、荧光光谱法、等温滴定量热法、凝胶阻滞法、微量热泳动法以及流体动力分散技术等.这些新技术方法的持续进步将极大地推动核酸适配体在筛选和应用等领域的技术方法创新,为核酸适配体的进一步商品化铺平道路.Nucleic acid aptamers which can be synthesized in vitro have attracted more and more attention in the field of life and health,and their binding affinity with targets is one of the important performance indexes.There are many methods to evaluate the binding affinity of nucleic acid aptamers.We screen and classify the technical methods,and describe the basic principle,research progress,and bottleneck problems of each technical method in detail.Mature methods for evaluating the binding affinity of nucleic acid aptamers include surface plasmon resonance,biological layer interference,enzyme-linked aptamer assay,and quartz crystal microbalance.These techniques can stably and sensitively capture molecular binding signals,which significantly promote the research progress of nucleic acid aptamer technology.However,the traditional aptamer immobilization technology may have a certain impact on the molecular conformation,thus changing its actual binding affinity in the environment.In order to solve this problem,researchers are actively exploring new methods to evaluate the binding affinity of molecules without immobilization,such as circular dichroism,UV–visible absorption spectroscopy,fluorescence anisotropy,fluorescence spectroscopy,isothermal titrition calorimetry,gel retardation,microscale thermophoresis,and flow induced dispersion analysis.The continuous progress of these new technologies and methods will greatly promote the innovation of nucleic acid aptamers in the fields of screening and application,and pave the way for further commercialization of nucleic acid aptamers.

关 键 词：核酸适配体 结合亲和力 分类策略 评测方法 

分 类 号：O64[理学—物理化学]