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人脑脊液中溶酶体来源酶的稳定性

Stability of enzymes of lysosomal origin in human cerebrospinal fluid.

作者信息

Goi G, Fabi A, Lombardo A, Burlina A B, Tiby V, Visciani A, Malesani L, Tettamanti G

出版信息

Clin Chim Acta. 1987 Mar 16;163(2):215-24. doi: 10.1016/0009-8981(87)90025-8.

Abstract

The optimal assay conditions and the stability of the following enzymes of lysosomal origin in human cerebrospinal fluid (CSF) were studied: acid phosphatase, beta-D-N-acetylglucosaminidase, alpha-D-galactosidase, beta-D-galactosidase, alpha-D-glucosidase, beta-D-glucosidase, alpha-L-fucosidase, alpha-D-mannosidase, beta-D-glucuronidase. The microsomal alpha-D-mannosidase, pH 5.7, was used as a reference non-lysosomal glycohydrolase. All the examined enzymes, with the only exception of beta-D-glucuronidase, underwent a more or less rapid loss of activity upon CSF storage in the temperature range from 37 degrees C to -80 degrees C. Storage in liquid nitrogen (-196 degrees C) was the only condition in which full activity for all tested enzymes was maintained for at least 15 days. Addition of human serum albumin to CSF, immediately after withdrawal, had a double effect in favouring enzyme stabilization and causing enzyme activation in some cases, and enzyme inhibition in others. Using conditions warranting enzyme stability the fluorimetric methods for lysosomal enzymes determination in cerebrospinal fluid appear to be highly reproducible (CV less than 5%) and simple enough for routine use.

摘要

研究了人脑脊液(CSF)中以下溶酶体来源酶的最佳测定条件和稳定性:酸性磷酸酶、β-D-N-乙酰氨基葡萄糖苷酶、α-D-半乳糖苷酶、β-D-半乳糖苷酶、α-D-葡萄糖苷酶、β-D-葡萄糖苷酶、α-L-岩藻糖苷酶、α-D-甘露糖苷酶、β-D-葡萄糖醛酸酶。微粒体α-D-甘露糖苷酶(pH 5.7)用作参考非溶酶体糖水解酶。除β-D-葡萄糖醛酸酶外,所有检测的酶在CSF于37℃至-80℃温度范围内储存时,活性或多或少会迅速丧失。液氮(-196℃)储存是唯一能使所有测试酶的活性至少维持15天的条件。在抽取CSF后立即向其中添加人血清白蛋白,对酶的稳定化有双重作用,在某些情况下会导致酶激活,而在其他情况下会导致酶抑制。在保证酶稳定性的条件下,脑脊液中溶酶体酶的荧光测定方法似乎具有高度可重复性(变异系数小于5%)且足够简单,可用于常规检测。

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