The effects of medium pH and purification method on product rate of expression rhGM-CSF in recombinant E.coli
- VernacularTitle:培养基pH值与提纯方法对大肠杆菌表达rhGM-CSF产率的影响
- Author:
Yisheng CAO
;
Xifeng WANG
;
Manrong BIN
;
Bing WANG
- Publication Type:Journal Article
- From:
Chinese Journal of Biochemical Pharmaceutics
2001;22(2):78-79
- CountryChina
- Language:Chinese
-
Abstract:
Purpose The aim is to study the influence factors of recombinant E.coli cell growth,product expression and purification,in order to raise the product rate of rhGM-CSF.Methods Three pH media were planed to observe the effects of pH on E.coli cell growth and expression with the indexes of the cell density and expression level.The influences of various treatments on product rate were observed with the indexes of the volume and purity of inclusion bodies and the yield,purity,activity of product.Using SDS-PAGE method for analysis of purity.The product bioactivity was determined by using TF1 cell and MTT colorimetric method.Results The cell density in the medium pH 7.0 was 2.22 g/L;the product expression rate was 23%.It was two times higher than other pH media.The inclusion body prepared by bacteria without freezing store was 2.5 times volume than freezing store.During renatura-tion PEG 4000 could raise 0.8 times active product.The optimum conditions used rhGM-CSF product rate could raise 8 times.Concluston The product rate of rhGM-CSF expressed in E.coli was affected by many factors.The optimum condition for culture,expression and purification could raise quite a number of product rate.
