Preparation,purification and biological function of fusion protein of ovalbumin and HSP70-1ike protein 1
- VernacularTitle:HSP70L1与OVA融合蛋白的制备及其生物学功能的初步研究
- Author:
Feng YANG
;
Tao WAN
;
Xiang-Yang ZHOU
;
Yan-Feng WU
;
Nan LI
;
Guo-You CHEN
;
Ying-Ming JIANG
;
Xue-Tao CAO
;
- Publication Type:Journal Article
- Keywords:
heat shock protein;
fusion protein;
dendritic cell;
adjuvant
- From:
Chinese Journal of Cancer Biotherapy
1995;0(02):-
- CountryChina
- Language:Chinese
-
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.
