CAJ | 학술논문

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0.2 M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25 KD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.
건립Protein A자주법검측원병독,장원병독항체IH2포피후적Protein A자주여단백매K소화과적공독소서뇌조직균장액혼합,경면역침정완충액온화세탈잡단백,재경0.2 M감안산장목적단백세탈병차이용취병선알응효전영검측.통과취병선알응효전영검측출25 KD적목적단백대,경여정상미소화적원단백조대상비교,증실검측출원병독.Protein A자주구유고효특이적여항체IgG결합적특점,여다극륭항체혼합후가유효결합IgG.해방법교면역조직화학화Western-blot기술간편쾌속,차자주가이회수재사용,사성본저렴.
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.h 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 Priori,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.