B.-S. YipHuang, Sheng-Cih; Wang, Yu-Kuo; Huang, Wan-Ting; Kuo, Tsam-Ming; Li, Tien-Hsiung Thomas; Wu, Tung-KungSheng-Cih; Wang, Yu-Kuo; Huang, Wan-Ting; Kuo, Tsam-Ming; Li, Tien-Hsiung Thomas; Wu, Tung-KungHuang2019-11-212019-11-212015https://www.scopus.com/inward/record.uri?eid=2-s2.0-84928047550&doi=10.1111%2fcas.12623&partnerID=40&md5=65577814427b4de42274d6eb02700916https://scholars.lib.ntu.edu.tw/handle/123456789/432021We report on the preparation of a new type of immunotoxin by conjugation of an epidermal growth factor receptor (EGFR)-binding peptide and an R46E mutation of thermostable direct hemolysin from Grimontia hollisae, (Gh-TDHR46E/EB). The hybrid immunotoxin was purified to homogeneity and showed a single band with slight slower mobility than that of Gh-TDHR46E. Cytotoxicity assay of Gh-TDHR46E/EB on EGFR highly, moderately, low, and non-expressed cells, A431, MDA-MB-231, HeLa, and HEK293 cells, respectively, showed apparent cytotoxicity on A431 and MDA-MB-231 cells but not on HeLa or HEK293 cells. In contrast, no cytotoxicity was observed for these cells treated with either Gh-TDHR46E or EB alone, indicating enhanced cytotoxic efficacy of Gh-TDHR46E by the EGFR binding moiety. Further antitumor activity assay of Gh-TDHR46E/EB in a xenograft model of athymic nude mice showed obvious shrinkage of tumor size and degeneration, necrosis, and lesions of tumor tissues compared to the normal tissues. Therefore, the combination of Gh-TDHR46E with target affinity agents opens new possibilities for pharmacological treatment of cancers and potentiates the anticancer drug's effect. ? 2015 The Authors. Cancer Science published by Wiley Publishing Asia Pty Ltd on behalf of Japanese Cancer Association.Grimontia hollisae; Anticancer; Epidermal growth factor receptor; Immunotoxin; Thermostable direct hemolysin[SDGs]SDG3bacterial toxin; celecoxib; cytotoxic agent; Grimontia hollisae thermostable direct hemolysin R46E epidermal growth factor receptor binding peptide fusion protein; hemolysin; hybrid protein; immunotoxin; unclassified drug; antineoplastic agent; bacterial protein; bacterial toxin; epidermal growth factor receptor; hemolysin; hemolysin, Vibrio; immunotoxin; recombinant protein; thermostable direct hemolysin; animal cell; animal experiment; animal model; animal tissue; antineoplastic activity; Article; cancer cell line; cancer inhibition; cell viability; controlled study; cytotoxicity; cytotoxicity assay; drug efficacy; erythrocyte; female; Grimontia hollisae; HEK293 cell line; HeLa cell line; hemolysis; histology; human; human cell; IC50; in vitro study; in vivo study; mouse; mutation; nonhuman; priority journal; protein purification; tumor necrosis; tumor volume; tumor xenograft; Vibrio; animal; genetics; metabolism; nude mouse; tumor cell line; Vibrionaceae; Animals; Antineoplastic Agents; Bacterial Proteins; Bacterial Toxins; Cell Line, Tumor; Female; HEK293 Cells; HeLa Cells; Hemolysin Proteins; Humans; Immunotoxins; Mice; Mice, Nude; Receptor, Epidermal Growth Factor; Recombinant Proteins; Vibrionaceaejournal article10.1111/cas.12623