title: Screening and Identification of ssDNA Aptamer for Human GP73.
abstract: As one tumor marker of HCC, Golgi Protein 73 (GP73) is given more promise in the early diagnosis of HCC, and aptamers have been developed to compete with antibodies as biorecognition probes in different detection system. In this study, we utilized GP73 to screen specific ssDNA aptamers by SELEX technique. First, GP73 proteins were expressed and purified by prokaryotic expression system and Nickle ion affinity chromatography, respectively. At the same time, the immunogenicity of purified GP73 was confirmed by Western blotting. The enriched ssDNA library with high binding capacity for GP73 was obtained after ten rounds of SELEX. Then, thirty ssDNA aptamers were sequenced, in which two ssDNA aptamers with identical DNA sequence were confirmed, based on the alignment results, and designated as A10-2. Furthermore, the specific antibody could block the binding of A10-2 to GP73, and the specific binding of A10-2 to GP73 was also supported by the observation that several tumor cell lines exhibited variable expression level of GP73. Significantly, the identified aptamer A10-2 could distinguish normal and cancerous liver tissues. So, our results indicate that the aptamer A10-2 might be developed into one molecular probe to detect HCC from normal liver specimens.
authors.affiliation: Kingmed College of Laboratory Medicine, Guangzhou Medical University, Guangzhou 510182, China.
authors.forename: Jingchun
authors.initials: J
authors.lastname: Du
authors.affiliation: Kingmed College of Laboratory Medicine, Guangzhou Medical University, Guangzhou 510182, China.
authors.forename: Jianming
authors.initials: J
authors.lastname: Hong
authors.affiliation: Kingmed College of Laboratory Medicine, Guangzhou Medical University, Guangzhou 510182, China.
authors.forename: Chun
authors.initials: C
authors.lastname: Xu
authors.affiliation: Kingmed College of Laboratory Medicine, Guangzhou Medical University, Guangzhou 510182, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Cai
authors.affiliation: Department of Laboratory Medicine, The First Affiliated Hospital, Guangzhou Medical University, Guangzhou 510120, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Xiang
authors.affiliation: Kingmed College of Laboratory Medicine, Guangzhou Medical University, Guangzhou 510182, China.
authors.forename: Chengbo
authors.initials: C
authors.lastname: Zhou
authors.affiliation: Kingmed College of Laboratory Medicine, Guangzhou Medical University, Guangzhou 510182, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Xu
mesh_terms: D000906:Antibodies / Q000235:genetics; D052157:Aptamers, Nucleotide / Q000235:genetics; D014408:Biomarkers, Tumor / Q000235:genetics*; D006528:Carcinoma, Hepatocellular / Q000175:diagnosis / Q000235:genetics*; D045744:Cell Line, Tumor; D004277:DNA, Single-Stranded / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008113:Liver Neoplasms / Q000175:diagnosis / Q000235:genetics*; D008565:Membrane Proteins / Q000235:genetics*; D052156:SELEX Aptamer Technique
chemical_list: D000906:Antibodies; D052157:Aptamers, Nucleotide; D014408:Biomarkers, Tumor; D004277:DNA, Single-Stranded; C410824:GOLM1 protein, human; D008565:Membrane Proteins