Production of recombinant antibody against epithelial cell adhesion molecule (EpCAM) in a bacterial host and Evaluation anti-proliferative effect on breast cancer cell.

Background and Aim: Epithelial cell adhesion molecule (EpCAM) is a transmembrane cell surface glycoprotein that is highly expressed in epithelial cancers and at lower levels in normal epithelia. EpCAM plays an important role in the proliferation, migration and differentiation of cancer cells. Evidence for the role of EpCAM in the pathogenesis of cancers has led to the development of MAbs that selectively target this receptor. The use of single-chain fragment variable (scFv) antibodies has emerged as an alternative to the use of polyclonal and monoclonal antibodies for many types of cancer treatment. According to the role of EpCAM in tumor growth, aim of this study was production of anti-EpCAM Scfv into bacterial system and evaluation the anti-proliferative effect of anti-EpCAM on breast cancer cell.Methods: In order to production of recombinant antibody in bacterial host, a gene cassette was designed including light and heavy variable (VL,VH) chain .VL and VH were joined together by a linker to generate scFv. Gene cassette amplified through polymerase chain reaction (PCR). The PCR product was inserted into the expression vector (pET۳۲Ek/LIC). Expression cassette were transformed into Escherichia coli BL۲۱. Produced antibody was assayed by conventional methods and toxicity of scfv-anti EpCAM was evaluated by MTT assay.Results: The expression cassette of anti-EpCAM scFv construct was successfully cloned and expressed in E. coli host cells. The recombinant scFv showed ۳۱ kDa band on SDS-PAGE and Western blot. In vitro assessment of anti-EpCAM on cancer cells indicates proliferations of cancer cells were decreased following treatment with increasing concentrations of anti-EpCAM scFv.Conclusion: In conclusion, we demonstrated that administration of scFv anti-EpCAM could suppress tumor growth. Despite the critical role of antibodies in cancer treatment, cost of producing recombinant antibodies is an important issue. Bacterial hosts, such as Escherichia coli, are an alternative and economical expression system for production of non-glycosylated antibodies and their fragments.

کلیدواژه ها:

Breast cancer ، EpCAM ، scFv-Antibody.