ERYTHROPOIETIN: PURIFICATION, PROPERTIES, BIOGENESIS

这一建议的显著特点是：研究那些具有结构性的 可能与其直接相关的人促红细胞生成素的特性 造血祖细胞生物学活性的研究 活性中心的一级结构；低聚糖的测定 侧链结构；特定结构的分离和表征 贫血感染细胞中促红细胞生成素的细胞受体 Friend病毒的毒株；受体研究的扩展到正常 并将为此目的开发的方法应用于 研究人类细胞受体。一旦分离出促红细胞生成素 受体将被克隆，该受体的基因将用于测试 红系细胞分化假说。小鼠促红细胞生成素 也将被克隆并将DNA用于研究细胞的调节 促红细胞生成素基因的表达及其在肿瘤研究中的应用 促红细胞生成素的生物发生和分泌。另一个目标是开发一种 抗EPO单抗固相放射免疫分析 将分析的灵敏度提高了一个数量级。相同 将以抗EPO单抗为基础进行改良 免疫亲和纯化方法。

The salient features of this proposal are: The study of those structural features of human erythropoietin that may be directly involved with its biological activity on hemopoietic precursor cells; the study of the primary structure of the active site; the determination of oligosaccharide side chain structure; the isolation and characterization of specific cellular receptors for erythropoietin from cells infected with the anemia strain of the Friend virus; the extension of receptor studies to normal mouse cells and to apply the methods developed for that purpose to the study of human cellular receptors. Once isolated the erythropoietin receptor will be cloned and the gene for the receptor used to test a hypothesis regarding erythroid cell differentiation. Mouse erythropoietin will also be cloned and the DNA used for the study of the regulation of expression of the erythropoietin gene as well as for the study of erythropoietin biogenesis and secretion. Another aim is to develop a solid-phase radioimmunoassay using monoclonal anti-epo with the intention of improving the sensitivity of analyis by an order of magnitude. The same monoclonal anti-epo will be used as the basis for an improved immunoaffinity purification method.