ERYTHROPOIETIN: PURIFICATION PROPERTIES BIOGENESIS

该提案的突出特点是： 人促红细胞生成素的特征，可能直接涉及其 对造血前体细胞的生物活性; 活性部位的一级结构;寡糖的测定 侧链结构;特异性的分离和表征 来自贫血感染细胞的促红细胞生成素细胞受体 Friend病毒株;受体研究扩展到正常 小鼠细胞，并将为此目的开发的方法应用于 人类细胞受体的研究。 一旦分离出促红细胞生成素 受体将被克隆，受体的基因将被用来测试一种 关于红细胞分化的假说。 小鼠红细胞生成素 也将被克隆，DNA用于研究调节 促红细胞生成素基因的表达以及用于研究 促红细胞生成素的生物合成和分泌。 另一个目标是开发一个 使用单克隆抗EPO的固相放射免疫测定， 将分析的灵敏度提高了一个数量级。 相同的 单克隆抗EPO将被用作改进的 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.