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| Human CT55 Protein Prokaryotic Expression and Its Production of Monoclonal Antibody |
Jian-wei REN1,Jun LI2,Shang-ze LI3,**( ) |
1. Tibet University Medical College, Lasha 85000, China
2. ABclonal, Wuhan 430072, China
3. Zhongnan Hospital of Wuhan University, Wuhan 430071, China |
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Abstract In order to prepare the monoclonal antibody of Cancer testis 55 (CT55), the prokaryotic expression plasmid with human CT55 fragment was constructed first, and the plasmid was expressed in the Rosetta competent cell. The target protein was purified and immunized for 6 weeks young female BALB/c mice. According to the preparation method of the conventional monoclonal antibody, the spleen and myeloma cells (sp2/0) of mice was prepared to fuse together. The hybridoma cell lines were obtained by indirect ELISA method and 2 successive subclones, such as 3D8B7B12, 4C8E1C9 and 3D8C10G9. The results of ELISA and Western blot analysis showed that the screened cell lines could produce monoclonal antibodies, and the antibody could specifically recognize the prokaryotic expression and eukaryotic expression of CT55 protein. Specific antibodies can be used in immunofluorescence test, and fluorescence is mainly located at the edge of nucleus. The results showed that monoclonal antibodies against human CT55 protein were successfully prepared. The preparation of CT55 protein monoclonal antibody has laid the material foundation for the rapid etiological diagnosis of cancer of liver cancer, gastric cancer and colon cancer, and the structure and function of CT55 protein.
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Received: 23 June 2018
Published: 06 December 2018
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Corresponding Authors:
Shang-ze LI
E-mail: 342883635@qq.com
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