重组人GM－CSF／MCAF融合蛋白抗血清的制备与鉴定

The Preparation and Identification of Recombinant Human GM CSF/MCAF Fusion Protein Antibodies

重组人粒细胞巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）和人单核细胞趋化激活因子（ＭＣＡＦ）融合蛋白经ＳｅｐｈａｄｅｘＧ－７５和ＣＭ－ＳｅｐｈａｒｏｓｅＦＦ两步柱层析，获得了电泳纯的ＧＭ－ＣＳＦ／ＭＣＡＦ融合蛋白。为进一步研究其结构与功能，我们以纯化的该融合蛋白为抗原免疫家兔制备抗血清。ＤｏｔＥＬＩＳＡ和Ｗｅｓｔｅｒｎｂｌｏｔ试验表明，该抗血清效价高、特异性好，可分别与ＧＭ－ＣＳＦ／ＭＣＡＦ、ＧＭ－ＣＳＦ和ＭＣＡＦ发生反应。

Recombinant Human granulocyte macrophage colony stimulating factor(GM CSF) and human monocyte chemoattractant activating factor(MCAF) fusion protein was purified to be homogeneity by Sephadex G 75 gel filtration and CM Sepharose FF ion exchange chromatography.For studies of the fusion protein on structure and function,the purified protein was used to immunize rabbits for the preparation of antisera.The analysis of Dot ELISA and western blot showed that the antisera was effective and specific,and that the antisera could react with GM CSF/MCAF,GM CSF and MCAF,respectively.