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头孢噻肟钠人工抗原的合成与鉴定
Synthesis and Identification of Cefotaxime Immunogen
【摘要】 采用碳化二亚胺法将头孢噻肟钠(CTX)与牛血清白蛋白(BSA)以及卵清白蛋白(OVA)分别进行偶联,合成完全抗原(CTX-BSA,CTX-OVA),紫外吸收光谱(UV)和凝胶电泳(SDS-PAGE)进行鉴定并测定CTX-BSA的偶联比,结果显示:与CTX(235.0 nm)、BSA(278 nm)、OVA(232.0,278.0 nm)相比,CTX-BSA(213.0,275.5 nm),CTX-OVA(223.5,274.5 nm)特征吸收峰位移明显,CTX-BSA的偶联比为5.8∶1.CTX-BSA与BSA以及CTX-OVA与OVA的SDS-PEG图谱相比,完全抗原泳动速度滞后.以CTX-BSA免疫动物并用间接酶联免疫吸附法(ELISA)测定抗血清效价,血清效价≥1∶100000.以CTX对免疫血清进行间接竞争ELISA测定,CTX可有效抑制免疫血清效价.
【Abstract】 Cefotaxime Sodium(CTX)was linked to bovine serum albumin(BSA) and ovalbumin(OVA),and was used to synthesize complete antigen CTX-BSA,CTX-OVA by carbodiimide method respectively.Using ultraviolet absorption spectrum and gel electrophoresis to identify complete antigen and test coupling ratio.The results showed that the displacement of characteristic absorption peak shifted obviously,compared CTX-BSA(213.0 nm,275.5 nm),CTX-OVA(223.5 nm,274.5 nm) with CTX(235.0 nm),BSA(278 nm),OVA(232.0 nm,278.0 nm) according to ultraviolet absorption spectrum.The coupling ratio of CTX-BSA was 5.8∶1.The atlas of gel electrophoresis displayed that CTX-BSA moved slowly than BSA did,so did CTX-OVA.The serum valence immunized by CTX-BSA can reach to 1∶100000 by using indirect ELISA detection.The indirect competitive ELISA serum inhibition test illustrated that the serum characterized with the antigen-specific antibody response to CTX can clearly inhibit the serum titer.
【Key words】 cefotaxime sodium; ovalbumin; bovine serun albumin; carbodiimide method; complete antigen;
- 【文献出处】 鲁东大学学报(自然科学版) ,Ludong University Journal(Natural Science Edition) , 编辑部邮箱 ,2012年03期
- 【分类号】S859.84
- 【下载频次】221