Preparation Of Monoclonal Antibody Against Human IgG And Application In The Diagnosis Of TB-antibody IgG

Objective: To prepare the monoclonal antibody of mouse anti-human IgG by hybridoma technique, and was applied in the preparing of the colloidal gold strip. To establish a rapid, simple immunodiagnostic assay for diagnosis of TB-antibody IgG Methods: Balb/c mice were immunized for 4 times with IgG from human serum. The myeloma cell line SP2/0 were fused with the spleen cells of immunized Balb/c mouse by PEG 1000, the hybridoma cells were selected by HAT medium. Monoclonal antibodies were screened by indirect ELISA, the isotype and affinity of monoclonal antibodies anti-human IgG were identified by indirect ELISA. The colloidal gold was prepared with sodium citrate as reducing agent, the monoclonal antibody 1E1 was labeled with gold granule.Results: 11 positive clones anti-human IgG were obtained after limited dilute cloning, the eleven strains of hybridoma cells were designated as 5A8, 3C10, 5E3, 1E1, 1D1, 1E2, 1E3, 1G3, 2D2, 1C12, 5Bll.The result of the chromosome number of the 11 strains monoclonal antibodies was 96-107. The isotype of 5A8, 5E3, 5B11, 1D1, 1E2, 1E3, 1G3, 3C10was IgG1, the isotype of 1E1 was IgG2a, the isotype of 2D2 was IgG2b,the isotype of 1C12 was IgM. The affinity of 5A8, 3C10, 5E3, 1E1 can achieve 10~7. The titer of 5A8, 3C10, 5E3, 1E1 was 3.9×10~7. The results of strip to diagnose positive and negative solution were good.Conclusions: McAbs against human IgG were successfully obtained and could be used to establish colloidal gold strip for diagnosis of TB immunoglobulin IgG.