Construction and expression of an anti-CD71 mouse/human chimeric antibody (D2C)
- VernacularTitle:抗CD71人-鼠嵌合抗体(D2C)的构建和表达
- Author:
Shuo WANG
;
Lin JIANG
;
Qing YE
- Publication Type:Journal Article
- Keywords:
CD71;
Human/mouse chimerical antibody;
Construction;
Expression
- From:
Chinese Journal of Immunology
2001;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct the expression vectors of VH and VL from an anti -CD71 monoclonal antibody (line7579) , and express an anti-CD71 mouse/human chimeric antibody (D2C) in vitro.Methods:The variable regions of heavy and light chains from an anti -CD71 monoclonal antibody (line7579) cloned in the pGEM-T vectors were constructed respectively into the expression vectors,p?-Expr(including human IgG1 constant region) and p?-Expr(including human Ig? constant region).The expression plasmid vectors (Chi7579-p?-Expr and Chi7579- p?-Expr) ,confirmed by a clone-PCR and a further sequence analysis, were co-transfected by electroporation into the non-Ig-producing murine hybridomas SP2/0.Results:Screening for transfectomas secreting chimeric antibody was done by cross sandwich ELISA, which identified the transfectomas to secrete a chimeric antibody (D2C) with the human heavy constant region(C?) and light constant region(C?). FCM analysis of culture supernatants of the transfectomas demonstrated that the binding capacity of the chimeric Ab (D2C) to the antigen (CD71) was retained.Conclusion:The construction of the V H and V L expression vectors and the expression of an anti-CD71 mouse/human chimeric antibody (D2C) were accomplished successfully.
