Preparation of human scFv antibody against nasopharyngeal carcinoma and identification of its specificity

Abstract

Conclusion: The selected scFv antibody could specifically recognize and target nasopharyngeal carcinoma (NPC), and could be applied to clinical diagnosis and therapy. Objective: The aim was to construct and screen fully human anti-NPC single chain Fv fusion phage libraries, and to identify the specificity of the scFv antibody. Methods: Peripheral blood mononuclear cells of patients with NPC were immunized in vitro by NPC cells and transformed by Epstein-Barr virus. The total RNAwas used to construct the scFv libraries. By means of ELISA and immunochemistry, the positively bound scFv was selected and identified. The positive scFv was fused to EGFP, and was then expressed in E. coli strain BL21 (DE3) and purified. Furthermore, we observed the binding bioactivity. Results: The fusion protein has the biological activity of binding the NPC cells and emitting green fluorescence. In targeting experiments in vivo, the results showed that the fusion protein can successfully target the NPC.

Keywords::

• Prokaryotic expression
• binding bioactivity

Acknowledgments

This study was supported by National High Technology Research and Development Program of China (863 Program) (no. 2007AA021809 and no. 2007AA021811), and the Major State Basic Research Delelopment Program (973) (2010CB833605).

Declaration of interest: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.