Construction, Expression and Internalization Study of Human Anti-HBsAg Single Chain Antibody/EGFP Fusion Proteins Containing Arg9
- VernacularTitle:含Arg9的人抗HBsAg单链抗体／EGFP融合蛋白基因的构建、表达和内化作用的研究
- Author:
Qian XUE
;
Wei-Hong WEN
;
Yan-Ling MENG
;
Cai-Fang XUE
;
Yong ZHANG
;
Wei ZHANG
;
Tao WANG
;
Lin-Tao JIA
;
An-Gang YANG
- Publication Type:Journal Article
- Keywords:
Hepatitis B HBsAg ScFv Arg9
- From:
China Biotechnology
2006;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To construct, express and purify ScFvl4/EGFP fusion proteins which containing Arg9, and to study their binding activities and internalization functions. Methods: Arg9 gene was recombined into 5' terminal, 3' terminal of ScFv/EGFP gene and between them respectively before they were cloned into the expression vector pET32a. After induced in E. coli BL21 and purified, their binding activities and internalization were respectively analyzed by indirect ELISA and indirect immunofluorescence analysis. Results: DNA sequencing and restriction endonuclease digestion proved that the four fusion genes were correctly constructed. SDS-PAGE analysis and Western blot showed that they were successfully expressed and purified. Indirect ELISA confirmed that the expressed products had antigen specific binding activities. Indirect immunofluorescence analysis revealed the fusion protein containing Arg9 at its N terminal had much better internalization function, but never internalized into the cells which do not express HBsAg. Conclusion: The four fusion genes were constructed, expressed and purified successfully. The purified fusion proteins maintained the binding activities to HBsAg and the fusion protein containing Arg9 at its N terminal had much better internalization effect.
