title: Functional expression, characterization and application of the human S100A4 protein.
abstract: Preparations utilizing monoclonal antibodies against S100A4 provide useful tools for functional studies to investigate the clinical applications of the human S100A4 protein. In the present study, human S100A4 protein was expressed in Escherichia coli (E. coli) BL21 (DE3), successfully purified by diethylaminoethyl cellulose anion-exchange chromatography and identified by western blot analysis. Soluble S100A4 bioactivity was confirmed by Transwell migration and invasion assays in the human HeLa cell line. Monoclonal antibodies (mAbs) were generated utilizing the standard hybridoma method and were validated by enzyme-linked immunosorbent assay and western blot analysis. The antibody was then used to examine human gastric carcinoma specimens by immunohistochemistry. Recombinant S100A4 was functionally expressed in E. coli and promoted the migration and invasion of HeLa cells. Four hybridoma cell lines, which secreted mAbs specifically against human S100A4 protein, were obtained. One of the four mAbs, namely 2A12D10B2, recognized human S100A4 as indicated by immunohistochemical staining of human gastric carcinoma specimens and recombinant S100A4 was functionally expressed in E. coli. The mAbs of recombinant S100A4 were suitable for detecting S100A4 expression in human tissues and for investigating the subsequent clinical applications of the protein.
authors.affiliation: Performance Medicine Laboratory, Institute of Health and Environmental Medicine, Tianjin 300050, P.R. China.
authors.forename: Degang
authors.initials: D
authors.lastname: Wang
authors.affiliation: Performance Medicine Laboratory, Institute of Health and Environmental Medicine, Tianjin 300050, P.R. China.
authors.forename: Jianwei
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Performance Medicine Laboratory, Institute of Health and Environmental Medicine, Tianjin 300050, P.R. China.
authors.forename: Ziquan
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Performance Medicine Laboratory, Institute of Health and Environmental Medicine, Tianjin 300050, P.R. China.
authors.forename: Yunyun
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Performance Medicine Laboratory, Institute of Health and Environmental Medicine, Tianjin 300050, P.R. China.
authors.forename: Chuanxiang
authors.initials: C
authors.lastname: Xu
authors.affiliation: Performance Medicine Laboratory, Institute of Health and Environmental Medicine, Tianjin 300050, P.R. China.
authors.forename: Zhiqing
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Performance Medicine Laboratory, Institute of Health and Environmental Medicine, Tianjin 300050, P.R. China.
authors.forename: Xiaohua
authors.initials: X
authors.lastname: Liu
authors.affiliation: Performance Medicine Laboratory, Institute of Health and Environmental Medicine, Tianjin 300050, P.R. China.
authors.forename: Lei
authors.initials: L
authors.lastname: Wu
authors.affiliation: Performance Medicine Laboratory, Institute of Health and Environmental Medicine, Tianjin 300050, P.R. China.
authors.forename: Xuesi
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Performance Medicine Laboratory, Institute of Health and Environmental Medicine, Tianjin 300050, P.R. China.
authors.forename: Xiangyan
authors.initials: X
authors.lastname: Meng
authors.affiliation: Endoscopy Division, Tianjin Medical University Cancer Hospital and City Key Laboratory of Tianjin Cancer Center, Tianjin 300060, P.R. China.
authors.forename: Hua
authors.initials: H
authors.lastname: Li
authors.affiliation: Performance Medicine Laboratory, Institute of Health and Environmental Medicine, Tianjin 300050, P.R. China.
authors.forename: Hongtao
authors.initials: H
authors.lastname: Liu
authors.affiliation: Division of Zoological Systematics and Evolution, Institute of Zoology, Chinese Academy of Sciences, Chaoyang, Beijing 100101, P.R. China.
authors.forename: Zifeng
authors.initials: Z
authors.lastname: Jiang
authors.affiliation: Performance Medicine Laboratory, Institute of Health and Environmental Medicine, Tianjin 300050, P.R. China.
authors.forename: Tianhui
authors.initials: T
authors.lastname: Wang
mesh_terms: D006801:Humans; D000071999:S100 Calcium-Binding Protein A4; D009418:S100 Proteins / Q000235:genetics* / Q000378:metabolism*
chemical_list: D000071999:S100 Calcium-Binding Protein A4; D009418:S100 Proteins; C077591:S100A4 protein, human