两种果子狸血清IgG纯化方法的比较

Comparison of Two Methods to Purify IgG from Paguma larvate Serum

目的探讨一种具有简便快捷、高纯度、活性好的果子狸血清IgG纯化方法。方法比较Hitrap Protein A亲和层析和PAGE电泳两种纯化法对果子狸血清IgG的纯化,用PAGE还原电泳和Western-Blot法对IgG作纯度鉴定。结果对Hitrap Protein A纯化的果子狸血清IgG,其活性虽好,但纯度不高;而非还原PAGE电泳所纯化的果子狸血清IgG不但纯度高(>95%)、并具有较强的免疫活性。结论非还原PAGE电泳法纯化果子狸血清IgG,是一种具有高纯度、免疫活性强的纯化方法。

Objective To evaluate the procedures to purify IgG with simple and rapid,high purity and high biological activity from Paguma larvate serum. Methods The Paguma larvate serum IgG was purified by Hitrap Protein A affinity chromatography and 8% non-reduced SDS-PAGE. The purity was identified by 12% SDS-PAGE, and compared the method on purification of IgG from Paguma larvate serum. The purification efficiency of the two methods was compared by amino black staining and protein A-HRP blotting. Results The IgG purified from Protein A affinity chromatography showed a high protein A- HRP binding activity, but purity is lower than non-reduced SDS-PAGE and the IgG from non-reduced SDS-PAGE also showed same levels of protein A-HRP binding activity. Conclusion For Paguma larvate serum IgG, purification by 8% non-reduced SDS-PAGE is an ideal method.