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Construction, Expression, and Characterization of a Recombinant Immunotoxin Targeting EpCAM
Joint Authors
Lv, Minghua
Qiu, Feng
Li, Tingting
Sun, Yuanjie
Zhang, Chunmei
Zhu, Ping
Qi, Xiaokun
Wan, Jun
Yang, Kun
Zhang, Kui
Source
Issue
Vol. 2015, Issue 2015 (31 Dec. 2015), pp.1-11, 11 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2015-04-16
Country of Publication
Egypt
No. of Pages
11
Main Subjects
Abstract EN
Epithelial cell adhesion molecule (EpCAM) is a type I transmembrane glycoprotein overexpressed in human epithelioma but with relatively low expression in normal epithelial tissues.
To exploit this differential expression pattern for targeted cancer therapy, an EpCAM-targeted immunotoxin was developed and its antitumor activity was investigated in vitro.
An immunotoxin (scFv2A9-PE or APE) was constructed by genetically fusing a truncated form (PE38KDEL) of Pseudomonas aeruginosa exotoxin with an anti-EpCAM single-chain variable fragment (scFv).
ELISA and flow cytometry were performed to verify immunotoxin (scFv2A9-PE or APE) antigen-binding activity with EpCAM.
Cytotoxicity was measured by MTT assay.
Confocal microscopy was used to observe its cellular localization.
The results of ELISA and flow cytometry revealed that the immunotoxin efficiently recognized recombinant and natural EpCAM.
Its antigen-binding activity was relatively lower than 2A9.
MTT assay confirmed potent reduction in EpCAM-positive HHCC (human hepatocellular carcinoma) cell viability (IC50 50 pM).
Immunofluorescence revealed that the immunotoxin localized to endoplasmic reticulum 24 h later.
In conclusion, we described the development of an EpCAM-targeted immunotoxin with potent activity against tumor cells, which may lay the foundation for future development of therapeutic antibody for the treatment of EpCAM-positive tumors.
American Psychological Association (APA)
Lv, Minghua& Qiu, Feng& Li, Tingting& Sun, Yuanjie& Zhang, Chunmei& Zhu, Ping…[et al.]. 2015. Construction, Expression, and Characterization of a Recombinant Immunotoxin Targeting EpCAM. Mediators of Inflammation،Vol. 2015, no. 2015, pp.1-11.
https://search.emarefa.net/detail/BIM-1072365
Modern Language Association (MLA)
Lv, Minghua…[et al.]. Construction, Expression, and Characterization of a Recombinant Immunotoxin Targeting EpCAM. Mediators of Inflammation No. 2015 (2015), pp.1-11.
https://search.emarefa.net/detail/BIM-1072365
American Medical Association (AMA)
Lv, Minghua& Qiu, Feng& Li, Tingting& Sun, Yuanjie& Zhang, Chunmei& Zhu, Ping…[et al.]. Construction, Expression, and Characterization of a Recombinant Immunotoxin Targeting EpCAM. Mediators of Inflammation. 2015. Vol. 2015, no. 2015, pp.1-11.
https://search.emarefa.net/detail/BIM-1072365
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1072365