Cloning and expression of a human monoclonal anti-D Fab fragment in E. coli with the use of bacteriophage vector
- VernacularTitle:抗-D抗体Fab段基因的克隆及其在大肠杆菌中的表达
- Author:
Yongshui FU
;
Chaofu JIANG
;
Shunong LI
;
Lin XU
;
Guangqing YUAN
;
Kaiyuan CAO
- Publication Type:Journal Article
- Keywords:
Antibodies, anti-D;
Gene cloning;
Gene expression;
Escherichia coli
- From:
Chinese Journal of Pathophysiology
1986;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To clone and express a human monoclonal anti-D Fab fragment in E. coli and make benefits for the expression of the whole immunoglobulin molecules of anti-D. METHODS: The gene of anti-D Fab fragment was cloned into the phagemid vector pComb3. After analyzing by PCR and restriction site analysis, the recombinant was expressed in E. coli and the expressed protein was analyzed by SDS-PAGE and ELISA. RESULTS: The result of SDS-PAGE confirmed that E.coli expressed a 48 kD protein. The ELISA result demonstrated that the cell culture supernatant reacted with Rh+ group O human erythrocytes, but was not recognized by Rh-group O human erythrocytes. CONCLUSION: Expressed Fab fragment has the antigenic specificity for human erythrocytes.
