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1.
Chinese Journal of Microbiology and Immunology ; (12): 926-932, 2019.
Article in Chinese | WPRIM | ID: wpr-824811

ABSTRACT

Objective To prepare monoclonal antibodies against pneumonia serotype 33F polysac-charides (Pn33Fps) and hepatitis B virus ( HBV) surface proteins ( HBs) by using the conjugate of Pn33Fps and HBs as antigen. Methods The conjugate of Pn33Fps and HBs was used as antigen to immu-nize mice with different immune doses, different immune procedures and different immune sites. Mouse spleen cells with higher antibody level after immunization were isolated and fused with SP2/0 myeloma cells. The hybridoma cells were screened specifically with Pn33Fps or HBs to prepare corresponding monoclonal antibodies. Results Serum antibodies against Pn33Fps and HBs were induced by immunizing mice with the conjugate. Monoclonal cell lines capable of continuously expressing antibodies against Pn33Fps or HBs were obtained. It has been proved that the recovery rates of samples of Pn33Fps and HBs prepared in three bat-ches tested with ascites monoclonal antibodies prepared by these two monoclonal cell lines were between 95% and 105%. Conclusions Monoclonal antibodies against Pn33Fps and HBs could be prepared simul-taneously by immunizing mice with the conjugate of Pn33Fps and HBs and used for the quantitative detection of Pn33Fps and HBs.

2.
Chinese Journal of Microbiology and Immunology ; (12): 926-932, 2019.
Article in Chinese | WPRIM | ID: wpr-800138

ABSTRACT

Objective@#To prepare monoclonal antibodies against pneumonia serotype 33F polysaccharides (Pn33Fps) and hepatitis B virus (HBV) surface proteins (HBs) by using the conjugate of Pn33Fps and HBs as antigen.@*Methods@#The conjugate of Pn33Fps and HBs was used as antigen to immunize mice with different immune doses, different immune procedures and different immune sites. Mouse spleen cells with higher antibody level after immunization were isolated and fused with SP2/0 myeloma cells. The hybridoma cells were screened specifically with Pn33Fps or HBs to prepare corresponding monoclonal antibodies.@*Results@#Serum antibodies against Pn33Fps and HBs were induced by immunizing mice with the conjugate. Monoclonal cell lines capable of continuously expressing antibodies against Pn33Fps or HBs were obtained. It has been proved that the recovery rates of samples of Pn33Fps and HBs prepared in three batches tested with ascites monoclonal antibodies prepared by these two monoclonal cell lines were between 95% and 105%.@*Conclusions@#Monoclonal antibodies against Pn33Fps and HBs could be prepared simultaneously by immunizing mice with the conjugate of Pn33Fps and HBs and used for the quantitative detection of Pn33Fps and HBs.

3.
Chinese Journal of Immunology ; (12): 1514-1519, 2017.
Article in Chinese | WPRIM | ID: wpr-657704

ABSTRACT

Objective:Generation and identification of hybridoma to produce monoclonal antibody against MICA/B. Methods:SP2/0 cells were fused with murine splenocyte immunized with recombinant protein rMICA?012 to get hybridoma cell lines. The titer of the monoclonal antibody produced by 9B10 cell line was determined by ELISA and its specificity was tested by Western blot,Luminex mutiplex microsphere-based immunoassay and immunofluorescence assay. Results:Six hybridoma cell lines were selected by ELISA screening test. The minimum reaction concentration of mAb 9B10 was 0. 02 ng/μl,and the specificity of mAb 9B10 was determinated by Western blot,Luminex mutiplex microsphere-based immunoassay and immunofluorescence. Conclusion:The monoclonal antibody 9B10 was available to apply for the detection of MICA and MICB expression.

4.
Chinese Journal of Immunology ; (12): 1514-1519, 2017.
Article in Chinese | WPRIM | ID: wpr-660052

ABSTRACT

Objective:Generation and identification of hybridoma to produce monoclonal antibody against MICA/B. Methods:SP2/0 cells were fused with murine splenocyte immunized with recombinant protein rMICA?012 to get hybridoma cell lines. The titer of the monoclonal antibody produced by 9B10 cell line was determined by ELISA and its specificity was tested by Western blot,Luminex mutiplex microsphere-based immunoassay and immunofluorescence assay. Results:Six hybridoma cell lines were selected by ELISA screening test. The minimum reaction concentration of mAb 9B10 was 0. 02 ng/μl,and the specificity of mAb 9B10 was determinated by Western blot,Luminex mutiplex microsphere-based immunoassay and immunofluorescence. Conclusion:The monoclonal antibody 9B10 was available to apply for the detection of MICA and MICB expression.

5.
Academic Journal of Second Military Medical University ; (12)1985.
Article in Chinese | WPRIM | ID: wpr-550677

ABSTRACT

The mice B lymphocyte hybridoma cells were irradiated by 60Co?-rays with doses of 1, 2, 4, 6. 8Gy. A markedly dose-relation depression in cell survival rate, cell concentration and clone-forming rate was observed. D37 value of clone-forming rate was 8.26 Gy. Dose-relation depression was also observed in total production of monoclonal antibody of clones. But production of monoclonal antibody per clone was raised with radiation doses. These results indicate that ionization radiation depresses the survival activity of hybridoma cells, but stimulates the secretion of monoclonal antibody of survival cells.

6.
Chinese Journal of Immunology ; (12)1985.
Article in Chinese | WPRIM | ID: wpr-674638

ABSTRACT

Spleen cells from Balb/c mice immunized with HBsAg were fused with HGPRT deficiencyhybridoma cells that secret anti-horseradish peroxidase (HRP) monoclonal antibodies (McAb),and two bispecific McAb secreting triomas were obtainedit is reacted with HRP.

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