核酸适配体—靶标复合物解离常数测定方法研究进展

Research progress for measuring dissociation constant of aptamer-target complex

核酸适配体是通过体外指数富集配基系统进化技术筛选到的一小段单链DNA或RNA寡核苷酸片段,能够与靶标高亲和力、高选择性地结合。为了在实际检测中应用核酸适配体,必须深入了解核酸适配体—靶标的结合过程,如测定复合物的解离常数。本文综述了近年来有关核酸适配体—靶标复合物解离常数测定的方法。这些方法大致上分为两大类:一类是基于分离策略的测定方法,如透析法、超滤法、凝胶电泳法、毛细管电泳法和高效液相色谱法;另一类是基于均相策略的测定方法,如荧光强度法、荧光各向异性法、紫外可见分光光度法、圆二色谱法以及表面等离子体共振法等。本文对每种方法的原理、应用范围和特性进行了概括和比较,并对未来的发展趋势进行了展望。

Aptamers are single stranded DNA or RNA molecules,selected using in vitro techniques of systematic evolution of ligands by exponential enrichment,and can bind target molecules with high affinity and selectivity.In order to use aptamers in practical detection,it is necessary to make thorough understanding of aptamer-target binding,such as measuring dissociation constant of aptamer-target complex.This article reviewed methods for assessing aptamer-target binding in recent years.These methods were generally divided into two categories.One was separation based techniques such as dialysis,ultrafiltration,gel and capillary electrophoresis,and high performance liquid chromatography（HPLC）;the other was mixture based techniques such as fluorescence intensity and anisotropy,UV-Vis absorption and circular dichroism,surface plasmon resonance,and so on.This paper summarized and compared the principle,range of application and important features of each method and give prospects of development trend in the future.