小分子靶标的非固定化核酸适配体筛选技术研究进展  

Research advances in non-immobilized aptamer screening techniques for small-molecule targets

作　　者：胡洋洋 杨歌 屈锋[1] HU Yangyang;YANG Ge;QU Feng(School of Life Science,Beijing Institute of Technology,Beijing 100081,China;Institute of Medicinal Biotechnology,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100050,China)

机构地区：[1]北京理工大学生命学院,北京100081 [2]中国医学科学院北京协和医学院医药生物技术研究所,北京100050

出　　处：《色谱》2025年第4期297-308,共12页Chinese Journal of Chromatography

基　　金：国家自然科学基金(22274009,82204263,9921874010);北京市自然科学基金(7222310).

摘　　要：核酸适配体是通过指数富集配体系统进化(SELEX)技术获得的一段核糖核酸(RNA)或单链脱氧核糖核酸(ssDNA),可用于靶标的特异性识别。核酸适配体具有易于化学合成和修饰、高热稳定性、低毒性与低免疫原性等优势。小分子靶标的核酸适配体在新药研发、肿瘤治疗、疾病诊断、超快速灵敏检测、环境污染监测和毒品检测等领域具有广泛的应用前景。然而,小分子因结构简单、分子质量小且可供核酸结合的基团有限,导致靶向小分子的适配体结合不稳定,为适配体的筛选与传感器的开发带来了巨大挑战。高效的筛选技术是获取高性能适配体的关键,目前适用于小分子靶标的核酸适配体筛选技术主要分为3类:基于靶标固定的筛选技术、基于核酸库固定的筛选技术以及靶标非固定化的筛选技术。与其他两种筛选技术相比,靶标非固定化的筛选技术所需的筛选轮数更少且获得的适配体亲和力更高(通常在nmol/L水平)。本文总结了小分子靶标的非固定化核酸适配体筛选技术(包括氧化石墨烯(GO)-SELEX、毛细管电泳(CE)-SELEX和纳米金辅助(GNP)-SELEX)的原理、优缺点及应用进展。此外,本文总结了在适配体特异性评价中对照靶标的选择策略。Aptamers obtained through systematic evolution of ligands by exponential enrichment(SELEX)techniques are single stranded deoxyribonucleic acid(ssDNA)or RNA molecules capable of specifically recognizing target molecules.Such aptamers are easily chemically synthesized and modified,highly thermally stable,and are low toxicity and low immunogenicity.Aptamers that target small molecules have broad applications prospects for the development of new drugs,treating tumors,diagnosing diseases,monitoring environmental pollution,detecting drugs,and in ultrafast and sensitive detection applications.However,the simple structures and low molecular masses of small molecules,along with the limited number of binding groups available for interacting with nucleic acids lead to unstable aptamer-small molecule binding,which poses significant challenges for aptamer screening and sensor development.Efficient screening techniques are crucial for identifying aptamers with excellent performance characteristics.At present,the aptamer screening techniques suitable for small-molecule targets are mainly divided into three categories:target-immobilized-based screening technique,nucleic acid library-immobilized-based screening technique,and target-non-immobilized screening technique.Among them,target-non-immobilized screening technique require fewer screening rounds and result in aptamers with superior(typically nmol/L level)affinities.This paper summarized non-immobilized aptamer screening techniques for small-molecule targets,including principle,advantages,disadvantages and application progress associated with graphene oxide(GO)-SELEX,capillary electrophoresis(CE)-SELEX,and gold nanoparticle-assisted(GNP)-SELEX techniques.In addition,strategies for selecting control targets in aptamer-specific evaluation were summarized.

关 键 词：指数富集配体系统进化技术 小分子靶标 非固定化核酸适配体筛选技术 综述 

分 类 号：O658[理学—分析化学]