Production of single chain antibody-alkaline phosphatase fusion protein
- VernacularTitle:单链抗体-碱性磷酸酶融合蛋白的制备
- Author:
Jing XU
;
Junxia LIU
;
Xia LI
- Publication Type:Journal Article
- Keywords:
Single chain antibody;
Alkaline phosphatase;
Fusion protein
- From:
Chinese Journal of Immunology
2001;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:Making single chain antibody(scFv)-alkaline phosphatase(Ap)fusion protein.Methods:An expression vector pSTE2-C66-Ap was constructed by sequentially inserting the Ap coding region into plasmid pSTE2-8E5 and replacing the VH-VL fragment of 8E5 by VH-VL fragment of C66.The fusion protein scFvC66-Ap was expressed in E.coli.and analysed by SDS-PAGE and immunoblotting.Results:Have obtained the scFvC66-Ap fusion protein with a molecular weight of 75 kD.It bound a 60 kD molecule from KG1a cell proteins on immunoblotting membrane detected directly by Ap enzymatic activity.Conclusion:A method permits the production of scFv-Ap conjugates in E.coli.which can replace conventionally prepared Ap-labeled antibodies in immunoassays.
