摘要
小分子通常指分子量小于1 kD的天然化合物,它们在生物科学、医药卫生和环境科学等多领域有重要的研究价值。适配体(aptamers)作为一种新型的生物识别元件,可特异地与小分子等靶标以高亲和力结合。它们是由指数富集的配体系统进化(systematic evolution of ligands by exponential enrichment,SELEX)技术筛选而得的单链短核苷酸序列。与大分子靶标的适配体相比,小分子靶标的适配体常表现出筛选困难、亲和力低和特异性差等缺点。为了解决这些瓶颈问题,本文提供了一些小分子靶标适配体筛选与鉴定的策略。首先,设计小分子靶标适配体的筛选方案时,本文提出需将关注点放在文库设计、SELEX方法、扩增方法、筛选进程监测方法和适配体序列分析方法等关键环节或方法的选择上,依据实际情况选择或设计适宜的策略。其次,适配体的结构与亲和力直接影响着它们的应用,本文简要介绍了适配体二级结构和三级结构的分析方法以及常见的6种亲和力鉴定方法。最后,本文讨论了小分子靶标适配体筛选与鉴定中存在的一些问题,展望了未来发展前景。全文旨在为小分子靶标适配体筛选与鉴定的多个环节提供研究策略,为提高适配体筛选与鉴定效率提供参考。
Small molecules generally refer to natural compounds with a molecular weight less than 1 kD,which have significant research value in many fields such as biological science,pharmaceutical and health care,and environmental science,etc.Aptamers,as a new type of bio-recognition element,can bind specifically to small molecules and other targets with high affinity.They are single-stranded short nucleotide sequences selected by the systematic evolution of ligands by exponential enrichment(SELEX)technology.Compared with aptamers of large molecules,aptamers of small molecules often exhibit disadvantages such as difficult screening,low affinity,and poor specificity.To solve these bottleneck problems,the article provides some strategies of screening and identification of aptamers for small molecules.Firstly,when designing the screening plan for aptamers of small molecules,this article suggests focusing on key points such as library design,SELEX methods,amplification methods,screening process monitoring methods,and aptamer sequence analysis methods,then choosing or designing appropriate strategies based on the actual situation.Secondly,the structure and affinity of aptamers directly affect their applications.The article briefly introduces methods for analyzing the secondary and tertiary structures of aptamers and six common methods for affinity determination.Finally,the article discusses some problems existing in screening and identification of aptamers for small molecules and looks forward to future development prospects.The whole article aims to provide research strategies for multiple steps of screening and identification of aptamers for small molecules and offer references for improving the efficiency of aptamer screening and identification.
作者
张小娟
王梁华
ZHANG Xiao-Juan;WANG Liang-Hua(Medical experiment center,Shaoxing University,Shaoxing 312000,Zhejiang,China;Department of Biochemistry and Molecular Biology,College of Basic Medicine,Naval Medical University,Shanghai 200433,China)
出处
《中国生物化学与分子生物学报》
北大核心
2025年第11期1656-1667,共12页
Chinese Journal of Biochemistry and Molecular Biology
基金
绍兴文理学院校级科研项目(No.2022LG012)资助。
关键词
小分子
适配体
指数富集的配体系统进化
结构分析
亲和力鉴定
small molecule
aptamer
systematic evolution of ligands by exponential enrichment(SELEX)
structural analysis
affinity determination
作者简介
通讯作者:王梁华,Tel:13386271017,E-mail:lhwang@smmu.edu.cn。
