Preparation of fluorescent polyclonal antibody against human IgG and its application in the diagnosis of nephrosis
- VernacularTitle:抗人免疫球蛋白G多克隆荧光抗体的制备及在肾活检中的应用
- Author:
Chunguang DUAN
;
Yanfang LIU
;
Shoujing YANG
;
Caiyun WANG
;
Lu YU
- Publication Type:Journal Article
- Keywords:
Human immunoglobulin G;
Polyclonal;
Fluorescent antibody;
Pathologic diagnosis
- From:
Journal of Medical Postgraduates
2003;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To prepare polyclonal antibody against human immunoglobulin G(human IgG) and to use this antibody in the clinical diagnosis of nephrosis. Methods:New Zealand rabbit was immunized subcutaneously with human IgG.Enzyme linked immunosorbent assay(ELISA) was applied to reveal the titer of the prepared polyclonal antiserum against human IgG.Antiserum was purified with affinity chromatography,and the purified antibody was confirmed for its specificity by Western blot and immunohistochemical staining.The purified antbodies which have been indentified were fused with FITC(fluorescein isothiocyanate) and then analyzed by immunohistochemical staining.Finally the fused antibody was useed in the clinical diagnosis of nephrosis. Results: The titer of the obtained antiserum was up to(1∶128 000,) double agar diffusion test showed the titer of the antibody was 1∶32.By the method of affinity chromatography,we obtained purified antibody with the purity of 85%.ELISA,double agar diffusion and immunohistochemical staining tests showed that the specificity and titer of the antibody were not decreased sharply.The purified antibody fused with FITC also kept the specificity of the primitive antibody.When the FITC fused antibody was tried in nephrosis patients,it detected human IgG effectively. Conclusion:The polyclonal antibody can specifically recognize human IgG.This purified antibody fused with FITC can be used in the diagnosis of nephrosis.
