Nielsen K
Can J Vet Res. 1986 Apr;50(2):227-31.
A method for preparing purified IgA from bovine mammary secretions is described. Whey was initially fractionated by gel filtration and fractions containing IgA were pooled, concentrated and digested with pepsin. The digest was rechromatographed on the same type of gel twice and the resulting IgA preparation tested for purity by an enzyme immunoassay procedure. Five different preparations tested were found to contain no measureable IgM or IgG2 and 0.8% to 1.1% IgG1 on a weight basis. If colostral whey was digested with pepsin prior to chromatography, the IgA preparations contained 1.1% to 2% IgG1 and no measureable IgM and IgG2. The procedure provides a reasonably easy method of eliminating most of the contaminating IgG1 (dimeric) and allows preparation of quantities of IgA for immunochemical studies and standardization of serological techniques.
描述了一种从牛乳分泌物中制备纯化IgA的方法。乳清首先通过凝胶过滤进行分级分离,含有IgA的级分合并、浓缩并用胃蛋白酶消化。消化产物在同类型凝胶上重新进行两次色谱分析,所得IgA制剂通过酶免疫测定法检测纯度。所测试的五种不同制剂在重量基础上未检测到可测量的IgM或IgG2,且IgG1含量为0.8%至1.1%。如果初乳乳清在色谱分析之前用胃蛋白酶消化,则IgA制剂含有1.1%至2%的IgG1,且未检测到可测量的IgM和IgG2。该方法提供了一种相当简便的方法来去除大部分污染性IgG1(二聚体),并能够制备用于免疫化学研究和血清学技术标准化的IgA量。