消旋氧氟沙星抗体制备及其手性识别特性研究  

Study on monoclonal antibodies preparation and chiral recognition characteristics against racemic ofloxacin

作　　者：王伟华[1] 雷红涛[1] 孙远明[1] 王利英[1] 王保玲[1] 

机构地区：[1]华南农业大学食品学院/广东省食品质量安全重点实验室,广东广州510642

出　　处：《食品工业科技》2011年第9期198-202,共5页Science and Technology of Food Industry

基　　金：国家自然科学基金(20877029;30700663);广东省科技项目(cgzhzd0808;2009B040500002;2009A01010004)

摘　　要：采用活泼酯法(A)和直接EDC(1-乙基-3-(3-二甲氨丙基)碳二亚胺盐酸盐)法(D),合成两种包被抗原(OFL-A-OVA和OFL-D-OVA)和两种免疫抗原(OFL-A-BSA和OFL-D-BSA),然后分别用两种免疫抗原免疫小鼠获得与其对应的多克隆抗体,建立消旋氧氟沙星残留免疫检测方法。研究结果表明:紫外扫描图谱证明四种抗原均合成成功,OFL-A-BSA、OFL-D-BSA、OFL-A-OVA和OFL-D-OVA的偶联比分别为19.67、3.03、1.93和0.99。消旋氧氟沙星的间接竞争ELISA法的线性检测范围为16.35～444.10ng/mL,IC50为5.42ng/mL,最低检测限为6.23ng/mL。消旋氧氟沙星抗体与恩诺沙星、环丙沙星、加替沙星等其他同类药物的交叉反应率较低,表明消旋氧氟沙星抗体具有较高的特异性,且消旋氧氟沙星抗体对左旋氧氟沙星和右旋氧氟沙星的识别并非对等,对左旋的识别能力较大,左旋氧氟沙星对抗体的产生起主要作用。Active ester method and direct EDC method were used to synthesize two kinds of coating antigens（OFL-A-OVA and OFL-D-OVA）and two kinds of immunity antigens（OFL-A-BSA and OFL-D-BSA）.The objective of experiment was to product artificial antigen and polyclonal antibodies against racemic ofloxacin（OFL）,then by which balb/c mice were immunized and acquired polyclonal antibodies.Meanwhile,racemic ofloxacin residual immunity examination method was established.The results indicated that four artificial antigens synthesized successfully proved by ultraviolet scanning.Happen to carrier protein ratios of OFL-A-BSA,OFL-D-BSA,OFL-A-OVA and OFL-D-OVA were 19.67,3.03,1.93 and 0.99,respectively.Detection scope of racemic ofloxacin with indirect competitive ELISA was 16.35～444.10ng/mL,and IC50 value was 5.42ng/mL,and detection limit was 6.23ng/mL.The cross reaction rates between racemic ofloxacin antibody and enrofloxacin,ciprofloxacin,or gatifloxacin were low,which indicated racemic ofloxacin had high specificity.Meanwhile,the abilities were not on balance with greater recognition for L-ofloxacin than D-ofloxacin.L-Ofloxacin played a major role for production of antibody which had high affinity and specificity.

关 键 词：氧氟沙星 人工抗原 多克隆抗体 交叉反应率 手性识别 

分 类 号：Q939.91[生物学—微生物学]