Hou Jun, Zhang Jian, Guo Jing-Xia, Cheng Lin, Zhao Jing, Liu Jia, Xu Jun, Liu Ai-Xia, Song Yong-Ji, Mao Pan-Yong, Li Bo-An, Mao Yuan-Li
Medical University of PLA, Beijing 100853, China.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2013 Jun;27(3):231-3.
To prepare the monoclonal antibody (mAb) against tissue inhibitor of metalloproteinases I (TIMP-I) fusion protein.
TIMP-I gene was amplified from fibrotic human liver tissue by RT-PCR, then ligated with pQE31 to form recombinant plasmid pQE-TIMP-I and transformed into E. coli BL21. The protein induced by IPTG was purified by 6 x His-tag and used to immunize the BALB/c mice. The specific monoclonal antibodies (mAbs) were prepared by the cell fusion technique. Western Blot were used to detect specificity of mAbs.
The prokaryotic plasmid expressing the recombinant protein was constructed, and the TIMP-I recombinant protein was expressed and purified. Four hybridoma cell lines that secreted anti-TIMP-I mAbs were obtained. 3 of 4 mAbs were the IgG1 subtype. Western Blot indicated the mAbs showed specific combination with TIMP-I protein.
The TIMP-I recombinant protein is highly purified and has strong antigenicity. The anti- TIMP-I mAbs were prepared successfully.
