Preparation,characterization and preliminary application of monoclonal antibody against cystatin C
- VernacularTitle:人胱抑素C单克隆抗体的制备、鉴定及初步应用
- Author:
Tingmei CHEN
;
Jiafu FENG
;
Zhiguang TU
- Publication Type:Journal Article
- Keywords:
Cystatin C;
Monoclonal antibody;
PETIA
- From:
Chinese Journal of Immunology
2000;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To prepare monoclonal antibodies(McAb)against cystatin C(Cys C)and to establish the particle enhanced turbidimetric immunoassay(PETIA)for determining human serum Cys C.Methods:The prokaryotic expression vector pET32a(+)/Cys C was constructed and Cys C expression was induced.McAbs against Cys C were prepared with the hybridoma technique after mice were immunized with the purified recombinant protein.Then the McAbs were covalently attached to uniform microparticles,PETIA method for determination of human serum Cys C was established,and primary evaluation tests of methodology were performed.Results:Three hybridoma cell lines were obtained successfully,the secreted antibodies were isotype of IgG1,and Western blot confirmed that the antibodies reacted specifically to the Cys C protein.After one of the hybridoma cell lines was injected into mice abdominal cavity,the ascites abundant for McAb was obtained.The titer of the McAb against the purified protein was 1∶4?106.With the self-made McAb,PETIA for human serum Cys C was established.The primary evaluation tests of methodology revealed that self-established PETIA method had a satisfactory performance,which was equal to the import kit.Conclusion:The prepared McAb against Cys C is prepared,which could be used to establish PETIA for determining human serum Cys C.
