Preparation,purification and biological function of fusion protein of ovalbumin and HSP70-1ike protein 1 / 中国肿瘤生物治疗杂志

ABSTRACT

Objective:To prepare,purify the recombinant proteins of HSP70-like protein 1 (HSPTOL1) with a large fragment of chicken ovalbumin (OVA),and to investigate the bio-function of the fusion protein,providing a basis for fur- ther study of the effect and the mechanism of HSPTOL1 as an adjuvant.Methods:The vector containing HSP70L1 cDNA and large fragment of OVA was constructed.The expression of OVA-HSP70L1 fusion protein was induced and the products were purified from inclusion bodies by His-Trap metal chelation chromatography and DEAE ion-exchange chromatography. The bio-activity of the fusion protein was examined by detecting its ability to activate dendritic ceils and to promote the se- cretion of cytokines.Results:The vector was successfully constructed and the molecular weight of the fused OVA- HSPTOL1 protein (with a purity of over 95%) was 68 000.The fusion protein effectively promoted the maturation of den- dritic cells and the production of cytokines such as interleukin-12 and tumor necrosis factor-?.Conclusion:HSPTOL1 may be an effective adjuvant in the fusion protein and it may also promote antigen specific Thl type i mmol/Luno-respon- ses.