Nucleotides and aptamers containing boronic acid groups having biased binding to glycosylated proteins, and uses thereof

US9096856B2 · kind B2 · utility

Assignee

• Georgia State University Research Foundation · US

Inventors

• Binghe Wang · Marietta, US
• Minyong Li · Nanjing, CN
• Zhen Huang · Brooklyn, US
• Na Lin · Taipei, TW

Key dates

• Filing date: Jul 17, 2008
• Grant date: Aug 4, 2015
• Expiry date: Feb 26, 2032

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N2310/33
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

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.