Design, Expression, Purification, and Evaluation of Anti-HER2 scFv
نویسندگان
1 Biochemistry Department, Advanced Sciences and Technology Faculty, Medical Sciences Islamic Azad University, Tehran, Iran
2 Biology Department, Science and Research Branch, Islamic Azad University, Tehran, Iran
3 Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
4 Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
5 Biotechnology Department, Faculty of Advanced Sciences and Technology, Tehran Medical Sciences, Islamic Azad University
6 Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
doi
چکیده
Aims: The aim of the present study was to design and express an anti-HER2 single chain variable antibody fragment in E. coli BL21 (DE3) and evaluate its efficiency in recognition of HER2 protein. Materials & Methods: An approximately 746bp encoding gene fragment was cloned into pET28a and the recombinant protein was expressed in E. coli BL21 (DE3) strain. Following protein purification by affinity chromatography, western blotting and ELISA were used to evaluate the efficiency of anti-HER2 scFv against HER2 protein. Findings: E. coli can express the anti-HER2 scFv molecule possessing appropriate function and can detect this protein on the surface of breast cancer cells. Conclusion: This antibody fragment can be used in laboratory diagnostic methods for HER2 diagnostic approaches. Potential capability of this protein in immunohistochemical and imaging approaches against HER2 should be considered.
