Expression of recombinant human MUC1-MBP fusion protein in E coli DH5?,and its purification and detection of immune responses in mice
- VernacularTitle:重组人MUC1-MBP融合蛋白在大肠杆菌中的表达、纯化及其免疫活性测定
- Author:
Guixiang TAI
;
Jifeng ZHANG
;
Xun ZHU
- Publication Type:Journal Article
- Keywords:
MUC1-MBP;
Fusion protein;
CTL activity;
Tumor vaccine
- From:
Chinese Journal of Immunology
2000;0(11):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study MUC1 based cancer vaccine.Methods:MUC1 gene was inserted into pMAL-p2 vector and constructed recombinant pMAL-MUC1. MUC1-MBP fusion protein expression was induced by IPTG in E coli DH5? transformed by the recombinant pMAL-MUC1 .The fusion protein was analyzed by Western blot and purified by amylose affinity chromatography. The antiserum,T cell proliferation and CTL activity of spleen from C57 mice immunized by MUC1-MBP were determined respectively by ELISA,adding 3H-TdR and MTT.Results:Had successfully constructed pMAL-MUC1 expression vector,and purified MUC1-MBP and MBP. C57 mice immunized by MUC-MBP generated MUC1 specific antibody and CTL.The titer of polyclonal antibody to MUC1 was about 1∶5 760?3 221. CTL cytotoxicity to the MCF7 and lewis lung cancer cells respectively were at 47.7%?4.3% and 67.5%?6.5%.Conclusion:Human recombinant MUC1-MBP fusion protein activated T and B cell response in mice.The results suggested that the recombinant Muc1 may be used to develop protein vaccine against carcinoma.
