麻痹性贝毒单克隆抗体的制备和酶联免疫检测方法的建立

发布时间：2018-04-13 12:08

本文选题：麻痹性贝毒 + 石房蛤毒素　；参考：《中国免疫学杂志》2013年01期

【摘要】：目的:利用B淋巴细胞杂交瘤技术制备麻痹性贝毒(PSP)的单克隆抗体,以便建立快速、灵敏、有效的毒素检测方法。方法:采用甲醛法将半抗原石房蛤毒素(STX)与血蓝蛋白(KLH)偶联制备成完全抗原STX-KLH,免疫BALB/c小鼠,取其脾细胞与SP2/0骨髓瘤细胞融合,经筛选和克隆,HAT选择培养杂交瘤细胞,利用ELISA方法筛选出分泌抗STX-McAB的杂交瘤细胞株,并通过小鼠体内诱生腹水的方法获得单克隆抗体。结果:共获得4株能稳定分泌麻痹性贝毒抗体的阳性细胞株,建立了分析检测麻痹性贝毒的间接竞争酶联免疫方法。对PSP中STX组分的检出限为20 ng/ml,IC50为220 ng/ml;对GTX2/3的检出限为10 ng/ml,IC50为50 ng/ml。结论:所制备抗体具有高特异性和灵敏性,可用于研制高质量的国产快速检测麻痹性贝毒ELISA试剂盒。
[Abstract]:Objective: to establish a rapid, sensitive and effective method for the detection of paralytic shellfish venom PSPs by using B lymphocyte hybridoma technique.Methods: a complete antigen STX-KLH was prepared by coupling hapten STX with hemocyanin KLH by formaldehyde method. BALB/c mice were immunized with STX-KLH. The spleen cells were fused with SP2/0 myeloma cells. The hybridoma cells were selected by screening and cloning hat.Hybridoma cell lines secreting anti- were screened by ELISA method and monoclonal antibodies were obtained by inducing ascites in mice.Results: four positive cell lines were obtained to stably secrete antibodies against paralytic shellfish venom, and an indirect competitive enzyme-linked immunosorbent assay (Elisa) was established for the detection of paralytic shellfish virus.The detection limit of STX component in PSP is 20 ng / ml and that in GTX2/3 is 20 ng / ml and 50 ng / ml, respectively.Conclusion: the prepared antibody has high specificity and sensitivity and can be used to develop a high quality ELISA kit for rapid detection of paralytic shellfish virus.
【作者单位】：国家海洋环境监测中心;国家海洋环境监测中心赤潮与藻毒素实验室;
【基金】：国家高技术研究发展计划(863计划)项目(2006AA09Z163);国家高技术研究发展计划(863计划)项目(2007AA092001) 国家908专项(908-01-ZH3) 国家海洋局海洋赤潮灾害立体监测技术与应用重点实验室基金项目(MATHAB200918)资助
【分类号】：R392.4

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