Novel recombinant immunotoxin of EGFR specific nanobody fused with cucurmosin, construction and antitumor efficiency in vitro
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Cuimin Deng1,*, Jiani Xiong2,*, Xiaofan Gu1, Xiaoying Chen3, Shuifa Wu4, Zhe Wang1, Duanduan Wang5, Jinjin Tu1, Jieming Xie1
1Department of Pharmacology, Fujian Medical University, Fuzhou, Fujian, China
2Department of Oncology, Fujian Medical University Union Hospital, Fuzhou, Fujian, China
3Department of Experimental Teaching Center of Basic Medical Science, Fujian Medical University, Fuzhou, Fujian, China
4Department of Pharmacology, The 180th Hospital of PLA, Quanzhou, Fujian, China
5Department of Breast Surgery, Fujian Medical University Union Hospital, Fuzhou, Fujian, China
*These authors contributed equally to this work
Jieming Xie, email: [email protected]
Keywords: immunotoxins, nanobody, EGFR, cucurmosin
Received: November 16, 2016 Accepted: March 24, 2017 Published: April 07, 2017
Epidermal growth factor receptor (EGFR) overexpression is related to the increased aggressiveness, metastases, and poor prognosis in various cancers. In this study, we successfully constructed a new EGFR nanobody-based immunotoxin rE/CUS containing cucurmosin (CUS), The immunotoxin was expressed by prokaryotic system and we obtained a yield of 5 mg protein per liter expression medium. The percentage of it’s binding ability totumor cell lines A549, HepG2, SW116, which highly expressed EGFR was 55.6%, 79.6% and 97.1%, respectively, but SW620 was only 4.45%. rE/CUS has the ability to bind A549, HepG2, SW116 cells specifically, and the antigen binding capability was not affected because of extra part of CUS component. The rE/CUS significantly inhibited the cell viability against EGFR over expression tumor cell lines in a dose-and time-dependent manner. Moreover, rE/CUS also induced apoptosis of HepG2 and A549 mightily. Our results demonstrate that rE/CUS is a potential therapeutic strategy for treating EGFR-positive solid tumors.
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