Study on the construction and expression of anti-IL2R? genetically engineering an-tibody Fab
- VernacularTitle:抗人IL-2R?基因工程抗体Fab的构建和表达
- Author:
Jiali LI
;
Xun ZHU
- Publication Type:Journal Article
- Keywords:
IL-2R? Recombinant antibody Fab
- From:
Chinese Journal of Immunology
1985;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Abstract Objective: To construct and express anti-IL-2R? genetically engineering antibody Fab and identify its biological activity.Methods: PCR were used to construct cloning vector 5G1-pA22 of anti-IL-2R? antibody Fab,by digestion of vector 5G1-pA22, anti-IL-2R? an-tibody Fab gene fragment was obtained and then this fragment was coupled with digesting product of pET28b plasmid by EcoRI and HindIII toget a recombinant expression vector 5G1-pET28b, it was highly expressed in the E. coli BL21(DE3) in the form of inclusion bodies, compoing20% of the total protein. After the renaturation of antibody Fab, its binding activity was identified by ELISA and antibody competitive inhibitionassay in vitro. Results: Anti-IL-2R? antibody Fab was successfully constructed and highly expressed in the form of inclusion bodies, its yield wasup to 1 mg/ml.When applied to ELISA, antibody Fab exhibited good spectfic reactivity with rIL-2R, antibody competitive inhibition assay invitro indicated that antibody Fab can compete with IL-2 for binding IL-2R in activated T cell surface.As a result,the activation and proliferationof T cells were inhibited and inhibition percentage exceeded 90%. Conclusion: Had successfully constructed and expressed anti-IL-2R? geneti-cally engineering antibody. Fab,it had good biological ativity.
