title: In vitro selection and characterization of deoxyribonucleic acid aptamers against connective tissue growth factor.
abstract: Connective tissue growth factor (CTGF) is a secreted matricellular protein possessing complex biological functions. CTGF modulates a number of signaling pathways that are involved in cell adhesion, migration, angiogenesis, myofibroblast activation, extracellular matrix deposition and tissue remodeling. Aptamers are oligonucleic acid chains or polypeptides that bind with specific target molecules hence have the potential to be used in the detection and blockade of the targets. In this study, we selected CTGF-targeting DNA aptamers by using systematic evolution of ligands by exponential enrichment (SELEX). After 8 iterative rounds of selection, cloning, DNA sequencing and affinity determination, six aptamers with high affinities to CTGF were obtained. Among them, one (C-ap17P) binds with the N-terminal region (aa 1-190) and the other five (C-ap11, 12, 14, 15 and 18) bind with the C-terminal region (aa 191-350) of hCTGF specifically. The biological stability assay indicated that a representative aptamer, C-ap17P, could keep its integrity at a rather high level for at least 24 h in complete DMEM cell culture medium. These CTGF aptamers might be used as a easy and fast detection tool for CTGF and be developed as CTGF-specific inhibitors for both research works and clinical applications.
authors.affiliation: Department of Gastroenterology, The First Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, 710061, PR China. Electronic address: lish09doctor@sina.com.
authors.forename: Shuang
authors.initials: S
authors.lastname: Li
authors.affiliation: Research Center of Reproductive Medicine, School of Medicine, Xi'an Jiaotong University, Xi'an, 710061, PR China. Electronic address: hyw301@mail.xjtu.edu.cn.
authors.forename: Yongwei
authors.initials: Y
authors.lastname: Huo
authors.affiliation: Research Center of Reproductive Medicine, School of Medicine, Xi'an Jiaotong University, Xi'an, 710061, PR China. Electronic address: th301@mail.xjtu.edu.cn.
authors.forename: Hong
authors.initials: H
authors.lastname: Tian
authors.affiliation: Department of Gastroenterology, The First Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, 710061, PR China. Electronic address: zqn1988zqn@163.com.
authors.forename: Qiannan
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: Department of Gastroenterology, Shaanxi Provincial People's Hospital and the Third Affiliated Hospital, Xi'an Jiaotong University, Xi'an, 710068, PR China. Electronic address: lvyifei@sina.com.
authors.forename: Yifei
authors.initials: Y
authors.lastname: Lv
authors.affiliation: Department of Gastroenterology, The First Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, 710061, PR China; Department of Rheumatology, The First Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, 710061, PR China. Electronic address: haozhm66@126.com.
authors.forename: Zhiming
authors.initials: Z
authors.lastname: Hao
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry*; D001483:Base Sequence; D001665:Binding Sites; D015374:Biosensing Techniques; D055513:Connective Tissue Growth Factor / Q000032:analysis*; D006801:Humans; D052156:SELEX Aptamer Technique / Q000379:methods
chemical_list: D052157:Aptamers, Nucleotide; D055513:Connective Tissue Growth Factor
keywords: Aptamer; Connective tissue growth factor; Deoxyribonucleic acid; Dissociation constant; Systematic evolution of ligands by exponential enrichment