Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, Brazil.
Methods Mol Biol. 2022;2466:187-203. doi: 10.1007/978-1-0716-2176-9_13.
Aptamers are affinity-based oligonucleotide ligands raised against a target molecule, which might be of proteic or other nature. Aptamers are developed by using a reiterative in vitro selection procedure, named SELEX, in which the target is exposed to a combinatorial oligonucleotide combinatorial library. Target bound oligonucleotides are eluted, and PCR amplified followed by the next SELEX round. The process is repeated until no further increase in target binding affinity and specificity is achieved. Selected aptamers are identified and immobilized for protein purification. In view of their stability against denaturation and capability of renaturation, low costs of production, easiness of modification and stabilization, oligonucleotide aptamers are excellent tools as high-affinity ligands for applications of protein purification.
适体是针对靶分子产生的基于亲和力的寡核苷酸配体，靶分子可能是蛋白质或其他性质的分子。适体是通过使用一种称为 SELEX 的反复体外选择程序开发的，在该程序中，将靶暴露于组合的寡核苷酸组合文库中。与靶结合的寡核苷酸被洗脱，并进行 PCR 扩增，然后进行下一轮 SELEX。该过程重复进行，直到靶结合亲和力和特异性不再进一步提高为止。选择的适体被鉴定并固定用于蛋白质纯化。鉴于它们对变性的稳定性和复性能力、低生产成本、易于修饰和稳定化，寡核苷酸适体是作为高亲和力配体用于蛋白质纯化应用的极好工具。
