A novel single-chain Fv antibody for connective tissue growth factor against the differentiation of fibroblast into myofibroblast |
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| Authors: | Guoqiu Wu Xiyong Wang Xuepeng Deng Pengpeng Wu Xiulei Xue Xuejiao Yan Xihua Wang |
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| Affiliation: | (1) Center of Clinical Laboratory Medicine of Zhongda Hospital, Southeast University, Nanjing, 210009, China;(2) Medical School, Southeast University, Nanjing, 210009, China;(3) Biotechnology Center, Department of Life Science and Biotechnology, China Pharmaceutical University, Nanjing, 210009, China;(4) Department of Respiratory, Zhongda Hospital, Southeast University, Nanjing, 210009, China |
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| Abstract: | This study was aimed to investigate the effect of a single-chain fragment variable antibody of connective tissue growth factor (anti-CTGF scFv) against the differentiation of fibroblast into myofibroblast. The scFv antibody was firstly expressed in Escherichia coli cells and was then purified by affinity chromatography. The yield scFv protein reached a purity over 95% after purification. Immunoreactivity assay demonstrated that scFv possessed a special affinity toward CTGF. RT-PCR, western blot, and immunofluorescence experiments showed that increased expression of α-smooth muscle actin induced by TGF-β1 could be suppressed by this scFv antibody through inhibiting the phosphorylation of Akt. |
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