Preparation of sheep and cattle immunoglobulins with antibody activity by high-performance liquid chromatography.
Preparation of sheep and cattle immunoglobulins with antibody activity by high-performance liquid chromatography.
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高效液相色谱法制备具有抗体活性的羊、牛免疫球蛋白。
DOI：
10.1016/s0021-9673(01)87448-3
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
Randall C. Cutlip
中科院分区：
文献类型：
--
作者：
Schmerr Mj;Kathryn R. Goodwin;H. Lehmkuhl;Randall C. Cutlip
采用高效阴离子交换层析(HPIEC)和高效体积排阻色谱(HPSEC)对羊和牛血清中的IgM和Ig G进行纯化。用CM-Affi-Gel Blue柱层析法制备正常牛羊混合血清和感染两种不同病毒的绵羊血清，用于HPIEC。HPIEC后，将含有Ig G和Ig M的组份混合并浓缩，再用HPSEC进一步纯化。用免疫电泳法、蛋白A-琼脂糖亲和层析法和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法对HPIEC和HPSEC进行纯度鉴定。用抗牛IgG2的单抗检测IgG2和其他组分的IgG2污染。用中和免疫扩散法检测抗体效价。在含有IgG1和IgM的组份中，对这两种病毒都保持了抗体抗力。该高效液相色谱法是一种快速制备特异性免疫球蛋白的方法，可用于评价免疫试剂的纯度。
High-performance anion-exchange chromatography (HPIEC) and high-performance size-exclusion chromatography (HPSEC) wereused to purify serum IgM and IgG from sheep and cattle. Pooled serum from normal cattle and sheep and serum from sheep, infected with two different viruses, were prepared for HPIEC by chromatography on CM-Affi-Gel Blue. After HPIEC, fractions containing IgG and IgM were pooled and concentrated and further purified by HPSEC. The purity of fractions from HPIEC and HPSEC were evaluated by immunoelectrohoresis, protein-A-Sepharose affinity chromatography, and sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Monoclonal antibody specific for bovine IgG2 was used to assay for IgG2 and IgG2 contamination of other fractions. The antibody activity to ovine adenovirus 5 and to ovine progressive pneumonia virus was assayed by neutralization and immunodiffusion. Antibody antivity was retained against both viruses in the fractions containing IgG1 and IgM. This high-performance liquid chromatography procedure was a rapid preparative method to produce specific immunoglobulins and could be used to evaluate the purity of immuno-reagents.
影响因子：
2.9
作者：
Vanecek,G;Regnier,FE
通讯作者：
Regnier,FE