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Purification of the surfactant associate proteins A from human bronchoalveolar lavage by affinity chromatography on maltosyl-agarose and its identification
Author(s): 
Pages: 884-886
Year: Issue:  9
Journal: THE JOURNAL OF PRACTICAL MEDICINE

Keyword:  肺表面活性物质载体蛋白质类;
Abstract: 目的:提取及纯化人肺灌洗液中肺表面活性物质结合蛋白 A(SP-A).方法:由支气管肺泡灌洗液经 10 000 g离心 40 min获得的沉淀中提取 SP-A.沉淀溶解于 6M的尿素, SP-A蛋白重新复性后上清过 maltosyl-agarose柱进行亲和层析,用梯度 EDTA洗脱 SP-A,并过 surpose 6柱纯化.聚丙烯酰胺电泳和 Western blotting鉴定.结果:经亲和层析法可以从人肺灌洗液中提取出高纯度的 SP-A,聚丙烯酰胺电泳在 36 ku和 70 ku处可见清晰条带,在分子量 118 ku以上亦可看到 3条清晰条带,经 Western blotting鉴定均为特异性 SP-A蛋白.结论:采用 maltosyl-agarose亲和层析法可以从肺灌洗液中获得高纯度的 SP-A.为进一步研究 SP-A的功能和制备 SP-A单克隆抗体提供了充足的 SP-A来源.
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