[ Objective] To prepare the monoclonal antibody against chicken major histocompatibility complex class II molecules （MHC II）. [ Method ] The prokaryotic expression of the gene fragments of exons 2 -6 encoding alpha ...[ Objective] To prepare the monoclonal antibody against chicken major histocompatibility complex class II molecules （MHC II）. [ Method ] The prokaryotic expression of the gene fragments of exons 2 -6 encoding alpha chain of MHC II and exons 3 -6 encoding beta chain of MHC II were performed based on its protein sequences. After BALB/c mice were immunized with the purified fusion proteins, the mouse spleen cells were fused with mouse myeloma cells SP2/0. Then the positive hybridoma cells were screened and detected by indirect enzyme-linked immunosorbent assay （ELISA）. [ Result] One hybridoma cell strain secreting monoclonal antibody against alpha chain and two strains secreting monoclonal antibody against beta chain were obtained. These three hybridoma cell strains were named as MHC II alpha-4, MHC II betas-2 and MCH II betas-31, respectively. Their titers of ascites in indirect ELISA were 1 ： 256 000, 1 ： 256 000 and 1 ： 1 280 000, respectively. These antibodies could specifically recog- nize MHC II alpha chain or beta chain in western blotting. [ Conclusion] Three obtained hybridoma stains can stably produce the monoclonal antibody against chicken MHC class II molecules.展开更多
[ Objective] To prepare monoclonal antibodies against chicken immunoglobulin G （IgG） and improve the diagnostic level of specific antibodies in chickens. [ Method] Chicken IgG was isolated by saturated ammonium sulf...[ Objective] To prepare monoclonal antibodies against chicken immunoglobulin G （IgG） and improve the diagnostic level of specific antibodies in chickens. [ Method] Chicken IgG was isolated by saturated ammonium sulfate and purified by Sephadex G-200 column chromatography. Then the BALB/c mice were immunized by the chicken IgG, and the spleen cells were fused with mouse myeloma cells SP2/0. Finally, the positive hybridoma cells were screened and detected by indirect enzyme-linked immunosorbent assay （ELISA）. [ Result] Four hybridoma cell strains secre- ting monoclonal antibodies against chicken IgG were obtained and named as C44, C45, C67 and C68, and their ascites titers in indirect ELISA were 1 ： 640 000, 1 ： 320 000, 1 ： 640 000 and 1 ： 80 000, respectively. The monoclonal antibodies secreted by C44 and C45 could recognize light chains of chicken IgG and those secreted by C,67 and C68 could recognize heavy chains of chicken IgG. They all could not recognize IgG from duck, rabbit and swine. Additionally, the Ig type identification results showed that they all belonged to IgGl. [ Conclusion] Four cell strains of obtained hybridoma can stably produce the monoclonal antibodies against chicken IgG.展开更多
Bio-effects of survival and etching damage on cell surface and DNA strand breaks were investigated in the yeast saccharomyces cerevisiae after exposure by nitrogen ion with an energy below 40 keV. The result showed th...Bio-effects of survival and etching damage on cell surface and DNA strand breaks were investigated in the yeast saccharomyces cerevisiae after exposure by nitrogen ion with an energy below 40 keV. The result showed that 16% of trehalose provided definite protection for cells against vacuum stress compared with glycerol. In contrast to vacuum control, significant morpho- logical damage and DNA strand breaks were observed, in yeast cells bombarded with low-energy nitrogen, by scanning electron microscopy （SEM） and terminal deoxynucleotidyl transferase- mediated dUTP nick end labeling （TUNEL） immunofluorescence assays. Moreover, PI （propidium iodide） fluorescent staining indicated that cell integrity could be destroyed by ion irradiation. Cell damage eventually affected cell viability and free radicals were involved in cell damage as shown by DMSO （dimethyl sulfoxide） rescue experiment. Our primary experiments demonstrated that yeast cells can be used as an optional experimental model to study the biological effects of low energy ions and be applied to further investigate the mechanism（s） underlying the bio-effects of eukaryotic cells.展开更多
[ Objective] To compare the characteristics of monoclonal antibodies against infectious bursal disease virus （IBDV） prepared with two different immunogens, VP2 protein expressed by prokaryotic system and purified IB...[ Objective] To compare the characteristics of monoclonal antibodies against infectious bursal disease virus （IBDV） prepared with two different immunogens, VP2 protein expressed by prokaryotic system and purified IBDV. [Methed] IBDV VP2 gene was amplified by RT-PCR and expressed in a prokaryotic system. The recombinant protein was purified by affinity chromatography. IBDV was pudfied by ultracentrifugation. Balb/c mice were immunized with the purified recombinant protein and IBDV, respectively. The monoclonal antibodies were screened by ELISA. [ Result] Two cell lines secreting antibodies against IBDV VP2 protein were obtained, and their ELISA titers were 1：2 × 10^4. Four cell lines secreting antibodies against I BDV were produced, and their ELISA titers were 1：2× 10^6, 1：6 × 10^4, 1：1× 10^5 and 1：4 × 10^3, respectively. All monoclonal antibodies specifically bound to their own immunogen but did not react with other viruses or proteins. After 10 -20 passages, these cell lines still secreted antibodies stably. [Condusion] The monoclonal antibodies prepared with the recombinant IBDV VP2 protein or purified IBDV can induce immune resoonse in mice. and VP2 soecific monoclonal antibodies can be obtained with VP2 orotein expressed in the Drokarvotic system as immunoQen.展开更多
基金supported by the National Natural Science Foundation (30671537)
文摘[ Objective] To prepare the monoclonal antibody against chicken major histocompatibility complex class II molecules （MHC II）. [ Method ] The prokaryotic expression of the gene fragments of exons 2 -6 encoding alpha chain of MHC II and exons 3 -6 encoding beta chain of MHC II were performed based on its protein sequences. After BALB/c mice were immunized with the purified fusion proteins, the mouse spleen cells were fused with mouse myeloma cells SP2/0. Then the positive hybridoma cells were screened and detected by indirect enzyme-linked immunosorbent assay （ELISA）. [ Result] One hybridoma cell strain secreting monoclonal antibody against alpha chain and two strains secreting monoclonal antibody against beta chain were obtained. These three hybridoma cell strains were named as MHC II alpha-4, MHC II betas-2 and MCH II betas-31, respectively. Their titers of ascites in indirect ELISA were 1 ： 256 000, 1 ： 256 000 and 1 ： 1 280 000, respectively. These antibodies could specifically recog- nize MHC II alpha chain or beta chain in western blotting. [ Conclusion] Three obtained hybridoma stains can stably produce the monoclonal antibody against chicken MHC class II molecules.
基金supported by the National Natural Science Fund (30671537)
文摘[ Objective] To prepare monoclonal antibodies against chicken immunoglobulin G （IgG） and improve the diagnostic level of specific antibodies in chickens. [ Method] Chicken IgG was isolated by saturated ammonium sulfate and purified by Sephadex G-200 column chromatography. Then the BALB/c mice were immunized by the chicken IgG, and the spleen cells were fused with mouse myeloma cells SP2/0. Finally, the positive hybridoma cells were screened and detected by indirect enzyme-linked immunosorbent assay （ELISA）. [ Result] Four hybridoma cell strains secre- ting monoclonal antibodies against chicken IgG were obtained and named as C44, C45, C67 and C68, and their ascites titers in indirect ELISA were 1 ： 640 000, 1 ： 320 000, 1 ： 640 000 and 1 ： 80 000, respectively. The monoclonal antibodies secreted by C44 and C45 could recognize light chains of chicken IgG and those secreted by C,67 and C68 could recognize heavy chains of chicken IgG. They all could not recognize IgG from duck, rabbit and swine. Additionally, the Ig type identification results showed that they all belonged to IgGl. [ Conclusion] Four cell strains of obtained hybridoma can stably produce the monoclonal antibodies against chicken IgG.
文摘Bio-effects of survival and etching damage on cell surface and DNA strand breaks were investigated in the yeast saccharomyces cerevisiae after exposure by nitrogen ion with an energy below 40 keV. The result showed that 16% of trehalose provided definite protection for cells against vacuum stress compared with glycerol. In contrast to vacuum control, significant morpho- logical damage and DNA strand breaks were observed, in yeast cells bombarded with low-energy nitrogen, by scanning electron microscopy （SEM） and terminal deoxynucleotidyl transferase- mediated dUTP nick end labeling （TUNEL） immunofluorescence assays. Moreover, PI （propidium iodide） fluorescent staining indicated that cell integrity could be destroyed by ion irradiation. Cell damage eventually affected cell viability and free radicals were involved in cell damage as shown by DMSO （dimethyl sulfoxide） rescue experiment. Our primary experiments demonstrated that yeast cells can be used as an optional experimental model to study the biological effects of low energy ions and be applied to further investigate the mechanism（s） underlying the bio-effects of eukaryotic cells.
文摘[ Objective] To compare the characteristics of monoclonal antibodies against infectious bursal disease virus （IBDV） prepared with two different immunogens, VP2 protein expressed by prokaryotic system and purified IBDV. [Methed] IBDV VP2 gene was amplified by RT-PCR and expressed in a prokaryotic system. The recombinant protein was purified by affinity chromatography. IBDV was pudfied by ultracentrifugation. Balb/c mice were immunized with the purified recombinant protein and IBDV, respectively. The monoclonal antibodies were screened by ELISA. [ Result] Two cell lines secreting antibodies against IBDV VP2 protein were obtained, and their ELISA titers were 1：2 × 10^4. Four cell lines secreting antibodies against I BDV were produced, and their ELISA titers were 1：2× 10^6, 1：6 × 10^4, 1：1× 10^5 and 1：4 × 10^3, respectively. All monoclonal antibodies specifically bound to their own immunogen but did not react with other viruses or proteins. After 10 -20 passages, these cell lines still secreted antibodies stably. [Condusion] The monoclonal antibodies prepared with the recombinant IBDV VP2 protein or purified IBDV can induce immune resoonse in mice. and VP2 soecific monoclonal antibodies can be obtained with VP2 orotein expressed in the Drokarvotic system as immunoQen.
