| 研究报告 |
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| Construction, expression and Internalization study of human anti-HBsAg single chain antibody/EGFP fusion proteins containing Arg9 |
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Abstract To construct，express and purify ScFv14/EGFP fusion proteins which containing Arg9,and to study their binding activities and internalization function. METHODS： Arg9 gene were recombined into 5'terminal、3'terminal of ScFv/EGFP gene or between them before they were cloned into the expression vector pET32a. After induced in E.coli.BL21 and purified, the binding activity and internalization of them were respectively analyzed heir binding activities and internalization function 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 activity. Indirect immunofluorescence analysis revealed fusion protein which 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 constructed, expressed and purified successfully. And the purified fusion proteins maintained the binding activity to HBsAg and fusion protein which containing Arg9 at its N terminal had internalization effect much better.
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Received: 18 January 2006
Published: 25 July 2006
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