title: Sequence-constructive SELEX: a new strategy for screening DNA aptamer binding to Globo H.
abstract: We proposed to use a novel stepwise sequence-constructive SELEX method to develop DNA aptamers that can recognize Globo H which is a tumor-associated carbohydrate antigen. A combinatorial synthetic library that consisted of DNA molecules with randomized regions of 15-bases was used as the starting library for the first SELEX procedure. The input DNA library for the second round of SELEX consisted of the extension of the 5' and 3'-ends with 7-bases that were randomized from four selected aptamers. The third round of SELEX was performed following the same procedures as described for the second round of SELEX. The experimental results indicate that the binding affinity of DNA aptamers to Globo H was enhanced when using the sequence-constructive SELEX approach. The selectivity of the DNA aptamers for related disaccharides, mannose derivatives, and Globo H analogs demonstrated the ability of the DNA aptamers to discriminate the presence of various glycans with different structures.
authors.affiliation: Genomics Research Center, Academia Sinica, 128 Academia Road, Sec. 2, Nankang, Taipei 11529, Taiwan.
authors.forename: Chin-Yu
authors.initials: CY
authors.lastname: Wang
authors.affiliation: Genomics Research Center, Academia Sinica, 128 Academia Road, Sec. 2, Nankang, Taipei 11529, Taiwan.
authors.forename: Chung-Yi
authors.initials: CY
authors.lastname: Wu
authors.affiliation: Genomics Research Center, Academia Sinica, 128 Academia Road, Sec. 2, Nankang, Taipei 11529, Taiwan.
authors.forename: Ting-Chun
authors.initials: TC
authors.lastname: Hung
authors.affiliation: Genomics Research Center, Academia Sinica, 128 Academia Road, Sec. 2, Nankang, Taipei 11529, Taiwan.
authors.forename: Chi-Huey
authors.initials: CH
authors.lastname: Wong
authors.affiliation: Genomics Research Center, Academia Sinica, 128 Academia Road, Sec. 2, Nankang, Taipei 11529, Taiwan. Electronic address: winschen@gate.sinica.edu.tw.
authors.forename: Chung-Hsuan
authors.initials: CH
authors.lastname: Chen
mesh_terms: D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000737:chemistry* / Q000378:metabolism; D052157:Aptamers, Nucleotide / Q000096:biosynthesis* / Q000737:chemistry; D001483:Base Sequence; D001665:Binding Sites; D004187:Disaccharides / Q000737:chemistry; D015723:Gene Library; D006801:Humans; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D052156:SELEX Aptamer Technique / Q000379:methods*
chemical_list: D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D015295:Antigens, Tumor-Associated, Carbohydrate; D052157:Aptamers, Nucleotide; D004187:Disaccharides; C472765:Globo-H
keywords: DNA aptamer; Globo H; SELEX