Turecek Frantisek, Scott C Ronald, Gelb Michael H
Department of Chemistry, University of Washington, Seattle, USA.
Methods Mol Biol. 2007;359:143-57. doi: 10.1007/978-1-59745-255-7_10.
Tandem mass spectrometry has been used for determinations of enzyme activities in biological samples. Activities in rehydrated dried blood spots of lysosomal enzymes glucocerebrosidase, acid sphingomyelinase, galactocerebroside beta-galactosidase, acid-alpha-galactosidase, acid alpha-glucosidase, and alpha-D-iduronidase are measured simultaneously by multiple-reaction monitoring of ion dissociations from cations produced by electrospray ionization of enzymatic products. Simple and inexpensive assay protocols are described that are readily adopted for handling multiple samples in 96-well microtiter plates, employing simple separation steps, and using less than or equal to 3 micromol of synthetic or commercially available substrates, and less than 25 nmol of internal standards per analysis. The assays have the potential of being used for large-scale screening of newborns for the detection of inborn errors of metabolism.
串联质谱已用于测定生物样品中的酶活性。通过对酶促产物电喷雾电离产生的阳离子的离子解离进行多反应监测，同时测量溶酶体酶葡萄糖脑苷脂酶、酸性鞘磷脂酶、半乳糖脑苷脂β-半乳糖苷酶、酸性α-半乳糖苷酶、酸性α-葡萄糖苷酶和α-D-艾杜糖醛酸酶在复水干血斑中的活性。本文描述了简单且廉价的检测方案，该方案易于在96孔微量滴定板中处理多个样品，采用简单的分离步骤，每次分析使用小于或等于3微摩尔的合成或市售底物，以及少于25纳摩尔的内标。这些检测方法有潜力用于大规模筛查新生儿以检测先天性代谢缺陷。
