Protein A磁珠检测朊病毒方法的建立  

The foundation of Protein A magnetism to detect Prion assay

作　　者：李奇[1] 杨阳[1] 吕鹏[2] 宋博翠[1] 白宗瑞[1] 陈志宝[1] 

机构地区：[1]黑龙江八一农垦大学生命科学学院,大庆163319 [2]吉林大学中日联谊医院手术室科

出　　处：《黑龙江八一农垦大学学报》2009年第3期73-75,94,共4页journal of heilongjiang bayi agricultural university

摘　　要：建立Protein A磁珠法检测朊病毒,将朊病毒抗体IH2包被后的Protein A磁珠与蛋白酶K消化过的攻毒小鼠脑组织匀浆液混合,经免疫沉淀缓冲液温和洗脱杂蛋白,再经0.2M甘氨酸将目的蛋白洗脱并且利用聚丙酰胺凝胶电泳检测。通过聚丙酰胺凝胶电泳检测出25KD的目的蛋白带,经与正常未消化的朊蛋白条带相比较,证实检测出朊病毒。Protein A磁珠具有高效特异的与抗体IgG结合的特点,与多克隆抗体混合后可有效结合IgG。该方法较免疫组织化学和Western-blot技术简便快速,且磁珠可以回收再使用,使成本低廉。Objective The country need the fast and effective detection for Prion asthe high risk country where Scrapie and BSE may bresk out. The common assay of Western-Blot and Immunohistochemistry consume a long time and the associated experience is needed. Protein A magnetism has the feature of binding IgG differential.It could bind the IgG adequately,when mixing with polyclonal antibody.After intermixing with brain tissue protein digested by protein kinase, Prion could easily elute from Protein A magnetism.Interest protein band is 25KD through SDS-PAGE. When definiting the molecular weight of Prion,the result of detection is educed by SDS-PAGE electrophoresis. The assay is more handier and faster than common assay. The magnetism can reclaim and reuse, which reduce the cost.The assay of detection is a innovation and many branches could generally applicated it.

关 键 词：PROTEIN A磁珠 朊蛋白(PrPC) 朊病毒(PrPSC) 免疫沉淀 

分 类 号：S852.65[农业科学—基础兽医学]