抗热毒宁注射液栀子蛋白单克隆抗体的研制与鉴定(3)
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Development and identification of monoclonal antibodies of cape
jasmine proteins in Reduning injection
LI Fang， ZHOU Jian，ming， WANG Hong，mei， ZHOU Jun， BI Yu，an， WANG Zhen，zhong， XIAO Wei*
(Jiangsu Kanion Pharmaceutical Co.， Ltd.， State Key Laboratory of New Pharmaceutical Process for Traditional Chinese Medicine，Enterprises Academician Workstations in Jiangsu Province， Lianyungang 222001， China)
[Abstract]Liposoluble cape jasmine proteins were used to immunize BALB/C mice. Indirect ELISA was utilized to develop one monoclonal antibody by integrating SP2/0 cells and spleen cells from immunized BALB/C mice. The subclass of the monoclonal antibody was identified as IgG2b， with Kappa chain as its light chain. The ascite titer of 2H8 monoclonal antibody was 1∶204 080. Western，blot analysis proved that 2H8 reacted with cape jasmine proteins to identify specific liposoluble protein with molecuar weight of around 58.5 kDa. Dot，ELISA was established with 2H8 ascites as the primary antibody， showing the minimum detectable amount of 19.5 ng. This study lays a foundation for the development of protein kits of Reduning injection.
[Key words]Reduning injection； protein； monoclonal antibody； Dot，ELISA
doi：10.4268/cjcmm201301031, 百拇医药(李芳 周建明 王红梅 周军 毕宇安 王振中 萧伟)
Development and identification of monoclonal antibodies of cape
jasmine proteins in Reduning injection
LI Fang， ZHOU Jian，ming， WANG Hong，mei， ZHOU Jun， BI Yu，an， WANG Zhen，zhong， XIAO Wei*
(Jiangsu Kanion Pharmaceutical Co.， Ltd.， State Key Laboratory of New Pharmaceutical Process for Traditional Chinese Medicine，Enterprises Academician Workstations in Jiangsu Province， Lianyungang 222001， China)
[Abstract]Liposoluble cape jasmine proteins were used to immunize BALB/C mice. Indirect ELISA was utilized to develop one monoclonal antibody by integrating SP2/0 cells and spleen cells from immunized BALB/C mice. The subclass of the monoclonal antibody was identified as IgG2b， with Kappa chain as its light chain. The ascite titer of 2H8 monoclonal antibody was 1∶204 080. Western，blot analysis proved that 2H8 reacted with cape jasmine proteins to identify specific liposoluble protein with molecuar weight of around 58.5 kDa. Dot，ELISA was established with 2H8 ascites as the primary antibody， showing the minimum detectable amount of 19.5 ng. This study lays a foundation for the development of protein kits of Reduning injection.
[Key words]Reduning injection； protein； monoclonal antibody； Dot，ELISA
doi：10.4268/cjcmm201301031, 百拇医药(李芳 周建明 王红梅 周军 毕宇安 王振中 萧伟)