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| Preparation and in vitro Activity of Anti-EpCAM Immunotoxin |
LIU Yu-ping1,DENG Chang-ping2,MA Xing-yuan2,LIU Qiu-li1,BAO Wen1,ZHENG Wen-yun1,**( ) |
1 Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China
2 State Key Laboratory of Bioreactor Engineering, School of Biological Engineering, East China University of Science and Technology, Shanghai 200237, China |
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Abstract Objective: Epithelial cell adhesion molecule(EpCAM) is one of the specific and effective targets of anti-tumor therapy, because the expression of EpCAM is limited to the basolateral epithelial cells in normal cells and is concealed. However, EpCAM is overexpressed in a variety of epithelial cancers, including bladder cancer, and is in a state of easy binding. Since immunotoxins targeting EpCAM have not been applied to the treatment of bladder cancer, new treatments for bladder cancer will be explored. Methods: The immunotoxin was constructed by linking the single-chain antibody of EpCAM 4D5MOCB with toxin PE38KDEL by flexible peptide GGGGS at the molecular level for design and preparation, and expressed in Escherichia coli BL21 (DE3). The binding effect of the immunotoxin on bladder cancer cells and its inhibitory effect on the growth of bladder cancer cells were studied. Results: The quantity of 4D5MOCB-mediated immunotoxin selectively binding to positive cells is about 169 times that binding to negative cells. The immunotoxin could effectively inhibit the growth of 5637, SW780 and RT4 with an IC50 up to 0.5 pmol/L. At the same time, it could inhibit cell colony formation and cell migration and induce cell apoptosis. The immunotoxin did not bind to EpCAM-negative HeLa cells and had no inhibitory activity. Conclusion: The immunotoxin prepared in this study has good selectivity and can effectively inhibit the proliferation and metastasis of positive cancer cells. The preparation of immunotoxin is simpler than that of antibody-drug conjugates (ADCs) and it is more homogeneous than ADCs. This study also provides an experimental basis for the application of immunotoxin in the treatment of solid tumors.
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Received: 04 May 2023
Published: 02 November 2023
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