Purification of a new low molecular weight protein and its clinical application

一种新型低分子量蛋白的纯化及其临床应用

• 批准号: 02454499
• 负责人: KAWAI Tadashi
• 金额: $ 2.82万
• 依托单位: JICHI MEDICAL SCHOOL
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-02454499/
• 关键词: protein 1; ELISA; low molecular weight protein; prostate; Clare cell; renal tubular dysfunction; protein 1; 低分子蛋白; protoin1; Clara cell secretary protein; urine protein1; human clara cell 10kDa Protein; 慢性腎不全

The authors have isolated a new low molecular weight protein (LMWP) (protein 1) from urine, characterized its physicochemical properties, determine it partial amino acid sequences, and thus prepared a monoclonal antibody against this protein. After establishing an enzyme-linked immunosorbent assay (ELISA) for this protein, the significance of this protein in laboratory medicine has been extensively studied.1) Purification and characterization of a new LMWP (protein 1):An unknown protein detected on SDS-PAGE so far proved to be protein 1 after checking the reactivity of rough purified fraction with various antisera (antibody). The present project is thus modified to isolate protein 1 using the antibody as a probe. The established purification procedure comprised of ammonium sulfate precipitation, ConA affinity, gel-filtration, anti-retinol binding protein antibody coupled affinity, and an anion exchange column. Purity was confirmed by SDS-PAGE and its molecular weight was 14kDa and p1 was 4.7. Partial amino acid sequences were determined which proved to be identical with human lung Clara cell 10 kDa secretary protein.2) Preparation of a monoclonal antibody.3) Establishment of an ELISA:A reliable precise assay was initially developed using purified substance as standard and its monoclonal anbitody.4) Clinical application:Protein 1 is distributed in various body fluids, in particular, serum value ranged in normal adults ranged from 2 to 20 mug/l and urine value was varied with marked sex-differences being higher in male than in female. Another extensive studies finally spot another synthetic sites as prostate. Further studies on this protein was both fundamentally and clinically in progress.

作者已经从尿液中分离出一种新的低分子量蛋白（LMWP）（蛋白1），表征了其物理化学特性，确定了部分氨基酸序列，因此制备了针对该蛋白的单克隆抗体。 After establishing an enzyme-linked immunosorbent assay (ELISA) for this protein, the significance of this protein in laboratory medicine has been extensively studied.1) Purification and characterization of a new LMWP (protein 1):An unknown protein detected on SDS-PAGE so far proved to be protein 1 after checking the reactivity of rough purified fraction with various antisera (antibody).因此，使用抗体作为探针进行了修改以将蛋白1隔离。已建立的纯化程序包括硫酸铵沉淀，CONA亲和力，凝胶过滤，抗归因醇结合蛋白抗体偶联的亲和力和阴离子交换柱。 SDS-PAGE证实了纯度，其分子量为14kDa，P1为4.7。确定了部分氨基酸序列，该序列与人类肺Clara细胞10 kDa秘书蛋白质相同。2）制备单克隆抗体的制备。3）建立ELISA：最初使用纯化的药物作为标准的及其单核的临床申请中的纯化剂量，在各种体内分布在各种体内。范围为2至20 mug/l，尿值的变化变化，男性的性别差异高于女性。另一项广泛的研究最终将另一个合成部位视为前列腺。对该蛋白质的进一步研究在根本上和临床上都是在进行的。

项目成果

R.Okutani: "Simple and high-yield purification of urine protein 1 using immuns affinity chromatography" J.chromatography. 577. 25-35 (1992)

R.Okutani：“使用免疫亲和层析对尿蛋白 1 进行简单且高产的纯化”J.层析。

Y.Itoh: "Increased urinary excretion of urine protein1 induad by L-Arginine monohydro chloride" Nephron. 62. 467-468 (1992)

Y.Itoh：“L-精氨酸单盐酸盐增加尿蛋白的尿排泄”肾单位。

y.Itoh et al.: "Increased urinary excretion of urine protein 1 by Lーarginine monohydrochloride" Nephron. (1992)

y.Itoh 等人：“盐酸拉精氨酸增加尿蛋白 1 的尿排泄”Nephron (1992)。

Y.Itoh,et al.: "Purification of urine protein 1 from urine: An initial study toward clinical application."

Y.Itoh 等人：“从尿液中纯化尿蛋白 1：临床应用的初步研究。”

Y. Itoh: "Increased urinary excretion of urine protein 1 induced by L-Arginine monohydrochloride" Nephron. 62. 467-468 (1992)

Y. Itoh：“L-精氨酸单盐酸盐诱导尿蛋白 1 的尿排泄增加”肾单位。