摘要
目的:用两种方法合成司帕沙星人工抗原,并制备相应的鼠源多克隆抗体。方法:用混合酸酐法和DCC法将半抗原SPFX与BSA偶联制备免疫原SPFX-BSA,通过DCC法制备人工包被原SPFX-OVA,采用紫外光扫描和聚丙烯酰胺凝胶电泳进行鉴定与分析。免疫小鼠后,采用间接ELISA法测定多抗血清效价,阻断ELISA,鉴定其抑制价和特异性。结果:UV和SDS-PAGE鉴定结果初步表明SPFX人工抗原成功合成;动物试验表明,4只小鼠多抗血清效价均在3.2×104以上,其中1号小鼠IC50值为163.87 ng/mL,与其他竞争物无明显交叉反应,抗体特异性良好。结论:两种偶联方法均能成功合成人工抗原,制备出鼠源抗SPFX多克隆抗体,其中混合酸酐法能明显提高完全抗原的合成和免疫效果。
Objective： This study was to synthesize artificial antigen of sparfloxacin with two methods and analyse its synthesis and characteristics. Methods： Artificial antigen SPFX-BSA were synthesized using the mixed anhydride reac- tion method and DCC method by linking carrier proteins BSA to SPFX and detection antigen SPFX-OVA were synthe- sized by DCC method, respectively synthesized immune antigen SPFX-BSA and detection antigen SPFX-OVA were identi- fied by UV and SDS-PAGE. BALB/c mice were immunized with SPFX-BSA. The mice polyclonal antiserum was identi- fied by ELISA and blocking ELISA. Results： It was successfully coupled when the wave moved to the right after BSA and SPFX was coupling as well as when the moving speed of BSA faster than SPFX in the SDS-PAGE electrophoresis. Four BALB/c mice indirect ELISA titers against SPFX were above 3.2×104 and the IC50 of No.1 mice was 163.87 ng/mL. Conclution： We can see that two kinds of Synthesis methods have an great effect on experimental result. This test has a better result by the mixed anhydride reaction method and SPFX polyclonal antiserum with high sensibility and specificity generated in this study may lay a foundation for the further SPFX monoclonal antibody preparation and the immunology fast detection methods.
出处
《中国食品学报》
EI
CAS
CSCD
北大核心
2014年第4期191-197,共7页
Journal of Chinese Institute Of Food Science and Technology
基金
农业部公益性行业(农业)科研专项(201003008)
