Volume 13, No 1, Feb 2003

ISSN: 1001-0602 
EISSN: 1748-7838 2018 
impact factor 17.848* 
(Clarivate Analytics, 2019)

Volume 13 Issue 1, February 2003: 35-48

ORIGINAL ARTICLES

Construction, expression and characterization of the engineered antibody against tumor surface antigen, p185c-erbB-2

Lian Sheng CHENG, Ai Ping LIU, Jia Hong YANG, Yan Qiu DONG, Liang Wei LI, Jing WANG, Chao Chen WANG, Jing LIU*

School of Life Science, University of Science and Technology of China, Hefei 230027, China Correspondence: Jing LIU(charlson@mail.ustc.edu.cn )

The c-erbB-2 proto-oncogene encodes a 185kDa protein p185, which belongs to epidermal growth factor receptor family. Amplification of this gene has been shown to correlate with poor clinical prognosis for certain cancer patients. The monoclonal antibody A21 which directed against p185 specifically inhibits proliferation of tumor cells overexpressing p185, hence allows it to be a candidate for targeted therapy. In order to overcome several drawbacks of murine MAb, we cloned its V_H and V_L genes and constructed the single-chain Fv (scFv) through a peptide linker. The recombinant scFvA21 was expressed in Escherichia coli and purified by the affinity column. Subsequently it was characterized by ELISA, Western blot, cell immunohistochemistry and FACS. All these assays showed the binding activity to extracellular domain (ECD) of p185. Based on those properties of scFvA21, we further constructed the scFv-Fc fusion molecule with a homodimer form and the recombinant product was expressed in mammalian cells. In a series of subsequent analysis this fusion protein showed identical antigen binding site and activity with the parent antibody. These anti-p185 engineered antibodies have promised to be further modified as a tumor targeting drugs, with a view of application in the diagnosis and treatment of human breast cancer.

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