李氏溶血素单克抗体制备及检测李氏溶血素的初步探索.pdfVIP

摘要 获得6株稳定分泌抗LLO单克隆抗体的杂交瘤细胞株，分别命名为I G7、ⅣB7、IVE9。 经亚类鉴定其中I 条；间接ELISA检测杂交瘤细胞培养上清抗体效价I 水单抗，经检测其效价为I：105，Westemblot检测该单克隆抗体具有很好的反应特异性；阻 断ELISA检测溶血素实验发现此单抗对天然李氏溶血素有较好的结合能力，不与葡萄球菌、 链球菌、丹毒杆菌所分泌的溶血索结合，说明所制备的抗重组LLO单克隆抗体对病原检测具 有高度的特异性。 盒奠定了基础。 关键词单核细胞增生性李斯特氏菌；李氏溶血素；单克隆抗体；鉴定；ELISA Of 0Monoclonal Listeriolysin Preparation And DetectionOfListeria Preliminary Abstract Listeria afoodbome whichcouldinfecthumanand monocytogenes(LMO)waspathogen animal．111ebacteriumcouldcausesevere as syndromes，such and case ratewas wassecreted fatality veryhigh．ListeriolysinO(LLO)whichby sepdcemia，the LMowasthevirulencefactorthatallowedtheLMOto fromthe escape primary vacuole．TheBALB／cwereimmunizedwith recombinantfusion 0which mice purified Listeriolysin was screenof expressedbyprokaryoticexpressionsystem 0McAbswas withnatural 0culturedinBHIculture anti-Listeriolysin performed Listeriolysin indirectELISA．SP2／0wasfusedwith cellsfromBALB／cmiceimmunize．The by spleen were cellsweresubclonedthreetimes selectedindirectELISA．The supernatants by positive by dilution．Sixstrains cellsline McAbsof O limiting hybridomasteadilysecreting wereobtainedandwerenamedI F5，IIC4，II E5，IIIG7，IVB7，IVE9，respectively． The ofMcAbssecretedI E5were were