| 摘要: |
| 以BSA-CAP-HS免疫Balb/C小鼠,用细胞融合技术筛选抗氯霉素单克隆抗体(CAP mAb)杂交瘤细胞,体内诱生腹水法制备CAP mAb,并鉴定其免疫学特性;依据胶体金免疫层析试验(GICA)原理,以CAP mAb 为金标抗体,研制CAP残留快速检测金标试纸(CAP-Strip),并对其性能进行测定。结果表明,筛选出了3株杂交瘤细胞,其中最好的4F9-B2株间接ELISA效价细胞培养上清为1:1 280,腹水为1:640 000,亲和常数(Ku)为 1.84×1010 L/mol,对CAP的半数抑制浓度(IC50)为0．88μg/L,与氯霉素琥珀酸钠的交叉反应(CR%)为150%,与其他酰胺醇类和抗菌素类药物无交叉反应;CAP-Strip的标准曲线呈典型的S型,符合4参数logit拟合曲线,目测检测限为0.5μg/L;其敏感性与竞争ELISA试剂盒相当,符合率为100％;可见CAP-Strip具有快速、敏感、特异、简便等特点,适合于CAP残留的快速检测,具有较高的推广应用价值。 |
| 关键词: 氯霉素 细胞融合 单克隆抗体 胶体金免疫层析试验 快速检测试纸 |
| DOI： |
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| 基金项目:国家“863”计划项目（2001AA249030） |
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| Filtration of hybridoma lines of monoclonal antibody and development of colloidal gold-labeled strip for rapid detection of chloramphenicol |
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| Abstract: |
| Balb/C mice were immunized with BSA-CAP-HS and hybridoma lines that secrete monoclonal antibody against chloramphenicol（CAP mAb）were filtered with cell fusion and the immunological traits of CAP mAb were characterized.Based on the principle of gold immunochromatography assay（GICA）,a stript for detection CAP（CAP-Strip）was developed with CAP mAb and its traits were tested.The results showed that three hybridoma lines were filtered and the best one was 4F9-B2,which secrete CAP mAb with indirect ELISA titers of 1∶1 280 in supernatant and 1∶640 000 in ascites.CAP mAb has a high affinity constant （Ka） with 1.84×1010 L/mol,a good sensitivity with an IC50 of 0.88 μg/L to CAP,150% cross-reactivity to CAP succinate sodium and little or no cress-reactivity to other compounds.The calibration curve of CAP-Strip was typical sigmoid curve fitted to the four parameter logistic.Its detection limit was 0.5μg/L and its sensitivity was as same as that of competitive ELISA-Kit and its coincidence rate was 100% compared with ELISA-Kit.The CAP-Strip possessed rapidity,sensitivity，specificity and briefness and was proved to be used in the rapid test of CAP residues in animal food. |
| Key words: chloramphenicol cell fusion monoclonal antibody gold immunochromatography assay rapid test strip |
