NUCLEOTIDES AND APTAMERS CONTAINING BORONIC ACID GROUPS HAVING BIASED BINDING TO GLYCOSYLATED PROTEINS, AND USES THEREOF
申请人:Wang Binghe
公开号:US20130184160A1
公开(公告)日:2013-07-18
The present disclosure encompasses oligonucleotide aptamers selectively binding a target glycosylated polypeptide or protein, and having biased affinity for the glycan through a boronic acid linked to a nucleosidic base of a nucleotide(s). The disclosure further encompasses methods for isolating an aptamer(s) selectively binding a target glycosylated polypeptide, where, from a population of randomized oligonucleotides that have at least one nucleotide having a boronic acid label linked to a base, is selected a first subpopulation of aptamers binding to the target glycosylated polypeptide or protein. This subpopulation is then amplified without using boronic acid-modified TTP, and amplification products not binding to a target glycosylated polypeptide or protein are selected. The second subpopulation of aptamers is then amplified using boronic acid-modified TTP to provide a population of boronic acid-modified aptamers capable of selectively binding to a glycosylation site of a target polypeptide or protein. Other aspects of the disclosure encompass methods for the use of the modified aptamers to detect glycosylated species of a polypeptide or protein.
本公开涵盖了寡核苷酸适配体,其通过与含有硼酸基团的核苷酸的核苷酸(s)相连,对靶向糖基化的多肽或蛋白质具有偏向性亲和力。本公开还涵盖了用于分离选择性结合目标糖基化多肽的适配体(s)的方法,其中从随机化寡核苷酸的群体中选择绑定目标糖基化多肽或蛋白质的第一亚群适配体。然后,在不使用硼酸修饰的TTP的情况下扩增该亚群,选择不与目标糖基化多肽或蛋白质结合的扩增产物。然后,使用硼酸修饰的TTP扩增第二亚群适配体,以提供能够选择性结合到目标多肽或蛋白质的糖基化位点的硼酸修饰适配体的群体。本公开的其他方面涵盖了使用修饰的适配体检测糖基化的多肽或蛋白质的物种的方法。