Manufacture and evaluation of a HER2-positive breast cancer immunotoxin 4D5Fv-PE25

Human epidermal growth factor receptor 2 (HER2) positive breast cancer is an aggressive subtype, accounting for around 20% of all breast cancers. The development of HER2-targeted therapy has substantially improved patient outcomes. Nevertheless, the increasing rate of side effects and resistance to...

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Bibliographic Details
Published in:	Microbial cell factories Vol. 22; no. 1; pp. 100 - 11
Main Authors:	Peng, Yanjie, Wu, Zhengli, Pang, Zheng, Zhang, Lin, Song, Dandan, Liu, Fang, Li, Yanhong, Lin, Tongjun
Format:	Journal Article
Language:	English
Published:	England BioMed Central Ltd 17-05-2023 BioMed Central BMC
Subjects:	Animals Antibodies Antibody-toxin conjugates Antimitotic agents Antineoplastic agents Apoptosis Biocompatibility Breast cancer Breast Neoplasms - drug therapy Breast Neoplasms - genetics Cancer therapies Caustic soda Cell Line, Tumor Chemical properties Chromatography Cytotoxicity Drug resistance E coli Efficiency ErbB-2 protein Escherichia coli - metabolism Evaluation FDA approval Female Fermentation Flow cytometry Freeze drying Growth factors HER2 Humans Hydrophobicity Immunotoxin Immunotoxins Immunotoxins - pharmacology Ion exchange Kinases Manufacture Mice Molecular weight Powders - therapeutic use Production processes Prokaryotic expression Protein synthesis Proteins Receptor, ErbB-2 - genetics Side effects Thymidine Toxicity Tumor cell lines Tumors Xenotransplantation China Evaluation Immunotoxin Breast cancer Prokaryotic expression Manufacture HER2
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Summary:	Human epidermal growth factor receptor 2 (HER2) positive breast cancer is an aggressive subtype, accounting for around 20% of all breast cancers. The development of HER2-targeted therapy has substantially improved patient outcomes. Nevertheless, the increasing rate of side effects and resistance to targeted drugs limit their efficacy in clinical practice. In this study, we designed and synthesized a new immunotoxin, 4D5Fv-PE25, which targets HER2-positive breast cancer, and evaluated its effectiveness in vitro and in vivo. The 4D5Fv-PE25 was expressed in high-density Escherichia coli (E. coli.) using the fermentor method and refined via hydrophobicity, ion exchange, and filtration chromatography, achieving a 56.06% recovery rate. Additionally, the semi-manufactured product with 96% purity was prepared into freeze-dried powder by the lyophilized process. Flow cytometry was used to detect the expression of HER2 in SK-BR-3, BT-474, MDA-MB-231, and MDA-MB-468 breast cancer cell lines. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) method was used for cytotoxicity assay, and the half-maximal inhibitory concentration (IC ) of 4D5Fv-PE25 lyophilized products to HER2-positive cell line SK-BR-3 was 12.53 ng/mL. The 4D5Fv-PE25 was injected into xenograft tumor mice via the tail vein on the 1st, 4th, and 8th day, it indicated that the growth of tumor volume was effectively inhibited for 24 days, although the 4D5Fv-PE25 was metabolized within 60 min by measuring the release of 3 H-Thymidine radiation. we succeeded in producing the 4D5Fv-PE25 freeze-dried powder using the prokaryotic expression method, and it could be employed as a potential drug for treating HER2-positive breast cancer.
ISSN:	1475-2859 1475-2859
DOI:	10.1186/s12934-023-02115-0