核酸适配体筛选的改良与优化策略

Improvement and Optimization of Aptamer Selection

核酸适配体以其类似抗体的功能及诸多优于抗体的特点引起人们的广泛关注,如何利用指数富集的配基系统进化技术(SELEX技术)来高效地筛选核酸适配体是其应用的关键。结合近些年核酸适配体的研究进展,为提高筛选效率及核酸适配体性能,针对适配体筛选三大不同环节所做的尝试及改良进行相关的阐述,包括初始随机寡核苷酸文库的设计与修饰、筛选过程所用方法及次级文库的制备、筛选过程后(Post-SELEX)核酸适配体性能的优化等;特别比较了筛选方法包括毛细管电泳SELEX、细胞SELEX、磁珠SELEX、加尾SELEX、微流控SELEX、微阵列SELEX和高通量SELEX,以及次级文库的制备方法诸如不对称PCR法、λ核酸外切酶法、磁珠法、不等长PCR法、综合性方法等;对核酸适配体发展面临的问题及发展前景进行探讨,以期为相关科研人员的研究提供参考。

Aptamers have gained extensive attentions due to their similar function as antibody and many superior characteristics,how touse systematic evolution of ligands by exponential enrichment（SELEX）technology to efficiently screen aptamers is the key to its application.Combined with the research progress of aptamers in recent years,the article introduces optimization strategies for increasing the screeningefficiency in three different stages of the screening process,including the design and modification of the initial random oligonucleotide librarybefore SELEX,the selection method and generation of ss DNA library during SELEX,and the optimization of aptamer performance afterSELEX（Post-SELEX）. Especially,the article compares many screening methods including capillary electrophoresis-SELEX,cell-SELEX,magnetic beads based-SELEX,tailored SELEX,microfluidic SELEX,microarray SELEX,and high-throughput SELE,as well as themethods for the generation of ss DNA such as asymmetric PCR,λ exonuclease digestion,magnetic beads-based separation,unequal lengthPCR,and some comprehensive methods. Also we explore the issues and prospects of the development of aptamers,providing references forrelated researchers.