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人尿中胰石蛋白的酶免疫测定及特性分析

Enzyme immunoassay and characterization of pancreatic stone proteins in human urine.

作者信息

Tatemichi N, Takahashi C, Hayakawa S, Hayakawa T, Shibata T, Kitagawa M, Sobajima H, Nakae Y

机构信息

Department of Clinical Chemistry, Maruko Pharmaceutical Co., Kasugai, Japan.

出版信息

J Clin Lab Anal. 1993;7(6):365-70. doi: 10.1002/jcla.1860070611.

Abstract

An enzyme immunoassay of pancreatic stone protein (PSP) in human urine was developed. Mean analytical recovery of pure PSP-S2-5 added to urine was 102.3% (SD 5.9%), and the precision of the assay was 2.0-2.7% within an assay and 2.5-2.9% between assays. In healthy volunteers (age 20-55 years), the mean value of the PSP concentration, expressed as ratios to urine creatinine, was 129 +/- 88 (mean +/- SD) micrograms/g without any differences for sex. Urine PSP correlated with urine N-acetylglucosaminidase (NAG) (r = 0.354). The molecular forms of immunoreactive PSP in urine were characterized by using cation exchange chromatography (Mono S), SDS-PAGE, N-terminal sequence, and enzyme immunoassay analysis. The urine PSP, eluted at the position corresponding to PSP-S2-5 on cation exchange chromatography, was converted to PSP-S1 by trypsin digestion. The difference in mobility on SDS-PAGE between urine PSP and PSP-S2-5 seems to be due to a glycosylated undecapeptide (N-terminal 1-11). The proposed method offers a sensitive, specific, and reproducible tool for laboratory analysis of human urine PSP levels.

摘要

已开发出一种检测人尿中胰石蛋白(PSP)的酶免疫分析法。添加到尿液中的纯PSP-S2-5的平均分析回收率为102.3%(标准差5.9%),该分析方法的精密度在批内为2.0 - 2.7%,批间为2.5 - 2.9%。在健康志愿者(年龄20 - 55岁)中,以与尿肌酐的比值表示的PSP浓度平均值为129±88(平均值±标准差)微克/克,性别之间无差异。尿PSP与尿N-乙酰氨基葡萄糖苷酶(NAG)相关(r = 0.354)。通过阳离子交换色谱法(Mono S)、SDS-PAGE、N端序列分析和酶免疫分析对尿中免疫反应性PSP的分子形式进行了表征。在阳离子交换色谱中对应于PSP-S2-5位置洗脱的尿PSP经胰蛋白酶消化后转化为PSP-S1。尿PSP与PSP-S2-5在SDS-PAGE上迁移率的差异似乎是由于一个糖基化的十一肽(N端1 - 11)。所提出的方法为实验室分析人尿PSP水平提供了一种灵敏、特异且可重复的工具。

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