Molecular Engineering Lab, Agency for Science Technology and Research, Singapore.
Biotechniques. 2011 Dec;51(6):413-6. doi: 10.2144/000113786.
Traditional methods for selecting aptamers require multiple rounds of selection and optimization in order to identify aptamers that bind with high affinity to their targets. Here we describe an assay that requires only one round of positive selection followed by high-throughput DNA sequencing and informatic analysis in order to select high-affinity aptamers. The assay is flexible, requires less hands on time, and can be used by laboratories with minimal expertise in aptamer biology to quickly select high-affinity aptamers to a target of interest. This assay has been utilized to successfully identify aptamers that bind to thrombin with dissociation constants in the nanomolar range.
传统的适体筛选方法需要经过多轮筛选和优化,才能鉴定出与靶标具有高亲和力的适体。在此,我们描述了一种仅需一轮正筛选,然后进行高通量 DNA 测序和信息学分析,即可筛选出高亲和力适体的方法。该方法具有灵活性,所需的人工操作时间更少,并且即使是适体生物学方面经验很少的实验室也可以使用,从而能够快速筛选出与目标靶标具有高亲和力的适体。该方法已成功地用于鉴定与凝血酶结合的具有纳摩尔级解离常数的适体。