摘要
目的在用乙二胺(ethanediamine,EDA)化牛血清白蛋白(BSA)为载体合成环丙沙星(ciprofloxacin,CIP)免疫原已制备了高特异性抗体(IC50达1 ng/ml)的基础上,尝试延长间隔臂长度,以己二胺(hexamethylenediamine,HMD)进一步修饰BSA合成免疫原制备抗血清,探索其对环丙沙星抗体灵敏度及特异性的影响。方法以己二胺修饰BSA,合成环丙沙星免疫原,免疫Balb/c小鼠制备多克隆抗体,同时以天然、乙二胺及己二胺化鸡卵清白蛋白(OVA)为载体合成包被原CIP-OVA、CIP-EDA-OVA和CIP-HMD-OVA,用间接竞争酶联免疫检测方法(ic-ELISA)对抗血清进行评价。结果获得了更高特异性的环丙沙星抗血清,IC50达0.1 ng/ml,抗体的交叉反应性也明显降低,且与CIP-EDA-OVA、CIP-HMD-OVA包被原相比,以CIP-OVA为包被原可显著提高环丙沙星ELISA灵敏度。结论改变抗原间隔臂长度对特异性抗体的产生有较大影响,且改变包被原间隔臂的结构可显著提高ELISA的灵敏度。
Based on the result that highly specific antibody can be obtained via immunogen prepared by ethylenediamine cationized BSA,this study is aimed to explore the effects of lengthening spacer arm between hapten and carriers on the sensitivity and specificity of antiserum against ciprofloxacin（CIP）.Hexamethylenediamine was employed to cationize BSA for preparing CIP immunogen,then polyclonal antibodies were raised in Balb/c mice immunized with CIP immunogen.Moreover coating antigens were synthesized using OVA and cationized OVA as carriers to screen antisera.The results showed that higher specificity and lower cross-reactivity antibody was raised with IC50 values of 0.1 ng/ml,which indicated that the length of spacer arm involved in immunogen has great effects on the raising of specific antibody.Furthermore,the heterology of space arm incorporated in coating antigen has great influence on the sensitivity of ELISA.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2011年第4期332-334,345,共4页
Immunological Journal
基金
国家高技术研究发展计划(863)资助项目(2007AA10Z438)
