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中国农学通报

中国农学通报 ›› 2009, Vol. 25 ›› Issue (15): 0-.

所属专题： 畜牧兽医

• 畜牧 动物医学 蚕 蜂 •    下一篇

奶牛初乳SIgA的提纯与电泳分析

梁靖坤,王继芳,曹荣峰   

  • 收稿日期:2009-03-20 修回日期:2009-04-02 出版日期:2009-08-05 发布日期:2009-08-05

The purification and electrophoretic analysis of SIgA in bovine colostrum

  • Received:2009-03-20 Revised:2009-04-02 Online:2009-08-05 Published:2009-08-05

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摘要： 为了建立提取奶牛初乳中SIgA的方法，本试验取分娩当日的奶牛乳汁，经10000 r/min离心10min，0.2mol/L（PH=6.0）PBS透析，上清液加0.06 mol/L硫酸锌处理后，调PH值为6.8-6.9，加入饱和硫酸铵，取沉淀溶于水，并在0.02mol/L（PH=8.0）PBS中透析至无NH4+，样品浓缩后上DEAE Sepharose (Fast Flow)柱，用0.02mol/L（PH=8.0）至0.3mol/L（PH=8.0）PBS梯度洗脱；收集峰蛋白上葡聚糖凝胶-200层析柱，用0.01mol/L（PH=7.4）PBS洗脱；收集出现蛋白峰的洗脱液，经SDS-PAGE电泳鉴定，为SIgA单体。经初步测定奶牛初乳中SIgA含量为3.47mg/ml。本试验已成功建立提取奶牛初乳中SIgA的方法。

Abstract: The experiment had establishmented the extraction method of SIgA in bovine colostrum. The milk of accouchement within one day was centrifuged 10 min by 10000r/min. After dialyzed by 0.2mol/L（PH=6.0）PBS, accommodated the supernatant fluid pH to 6.8-6.9 after disposal of 0.06 mol/L ZnSO4. Then added saturated ammonium sulfate to produce sedimentum, sedimentum dissolved in water. The supernatant fluid were dialyzed by 0.02mol/L（PH=8.0）PBS to no NH4+. Specimen was concentrated, then it was purified by DEAE Sepharose FF chromato bar, gradient elutioned by 02mol/L（PH=8.0）to reach 0.3mol/L（PH=8.0）PBS. Hump proteinum was collected, afterwards it was purified by Sephadex G-200. The elutriant was 0.01mol/L（PH=7.4）PBS. The elutriant appearance hump proteinum was collected, then it was appraised through electrophoresis of SDS-PAGE. The results of the electrophoresis showed that SIgA was metamer. The results of rough determination showed that the content of SIgA was 3.47mg/ml in bovine Colostrum. This experiment had established the method for extract SIgA in bovine colostrum.