Preparation of monoclonal antibody against recombinant GST and its application in purification of GST fusion protein
10.3321/j.issn:1007-8738.2001.04.023
- VernacularTitle:抗重组 GST单克隆抗体的制备及其在融合蛋白纯化中的应用
- Author:
Xinrui YAN
;
Yongli BAO
;
Xuebin DONG
;
Zhonghui LIU
- Publication Type:Journal Article
- From:
Journal of Cellular and Molecular Immunology
2001;17(4):365-367
- CountryChina
- Language:Chinese
-
Abstract:
Aim To prepare and characterize a monoclonal antibody against recombinant glutathione S-transferase(GST) for purifying GST fusion protein. Methods The GST-follistatin fusion protein was expressed by using a pGEX4T-1 expression vector in Escherichia coli BL21 and purified by glutathione-resin affinity column chromatography. Then female Balb/c mice were immunized with the GST-FS, The immunized splenocytes were fused with NS-1 hybridoma cells. Dreparation of the mAb was used by conventional hybridoma techniqal. The mAb purified by protein A, was culpled with Sepharose4B to purify further GST fusion protein by affinity chromatography. Results The SDS-PAGE showed that the GST fusion protein could be purified effctively by specific mAb affinity chromatography as same as by glutathione-resin affinity chromatography. Conclusion mAb affinity chromatography will be a ecnomical and useful method and it can be used for secondary purification of GST fusion protein following glutathione-resin affinity chromatography.
