Improved reagents for newborn screening of mucopolysaccharidosis types I, II, and VI by tandem mass spectrometry.
Improved reagents for newborn screening of mucopolysaccharidosis types I, II, and VI by tandem mass spectrometry.
复制标题
DOI：
10.1021/ac5004135
复制
发表时间：
2014-05-06
影响因子：
7.4
通讯作者：
Gelb, Michael H.
中科院分区：
文献类型：
--
作者：
Chennamaneni, Naveen Kumar;Kumar, Arun Babu;Barcenas, Mariana;Spacil, Zdenek;Scott, C. Ronald;Turecek, Frantisek;Gelb, Michael H.
串联质谱法用于新生儿筛查卡上干血斑中酶活性的多重和定量分析，已成为早期评估溶酶体贮积病的有力技术。在本文中，我们报告了用于新生儿I型、II型和VI型粘多糖样变性筛查的酶α-L-艾杜糖醛酸酶、艾杜糖醛酸-2-硫酸酯酶和N-乙酰半乳糖胺-4-硫酸酯酶底物的设计和工艺规模合成。该产品含有一个双酰胺单元，假设在气相中容易质子化，这提高了串联质谱法的检测灵敏度。该产品含有一个苯甲酰基，这提供了一个有用的网站，廉价的氘代，从而有利于制备内标物的酶的产品的准确定量。最后，试剂的设计考虑到易于合成，从而允许放大制备，以支持全球新生儿筛查溶酶体贮积病。新试剂提供了迄今为止报道的三种溶酶体酶的最灵敏的测定，如它们在使用干血斑作为酶源的反应中的性能所示。此外，测定信号与在不存在血液（背景）的情况下测量的信号的比率上级于所有先前报道的I型、II型和VI型粘多糖样变性测定。
Tandem mass spectrometry for the multiplex and quantitative analysis of enzyme activities in dried blood spots on newborn screening cards has emerged as a powerful technique for early assessment of lysosomal storage diseases. Here we report the design and process-scale synthesis of substrates for the enzymes α-l-iduronidase, iduronate-2-sulfatase, and N-acetylgalactosamine-4-sulfatase that are used for newborn screening of mucopolysaccharidosis types I, II, and VI. The products contain a bisamide unit that is hypothesized to readily protonate in the gas phase, which improves detection sensitivity by tandem mass spectrometry. The products contain a benzoyl group, which provides a useful site for inexpensive deuteration, thus facilitating the preparation of internal standards for the accurate quantification of enzymatic products. Finally, the reagents are designed with ease of synthesis in mind, thus permitting scale-up preparation to support worldwide newborn screening of lysosomal storage diseases. The new reagents provide the most sensitive assay for the three lysosomal enzymes reported to date as shown by their performance in reactions using dried blood spots as the enzyme source. Also, the ratio of assay signal to that measured in the absence of blood (background) is superior to all previously reported mucopolysaccharidosis types I, II, and VI assays.
登录
查看更多内容
影响因子：
7.4
作者：
Spacil, Zdenek;Elliott, Susan;Reeber, Steven L.;Gelb, Michael H.;Scott, C. Ronald;Turecek, Frantisek
通讯作者：
Turecek, Frantisek
影响因子：
1.3
作者：
Ombrone, Daniela;Malvagia, Sabrina;la Marca, Giancarlo
通讯作者：
la Marca, Giancarlo
影响因子：
9.3
作者：
Duffey, Trisha A.;Bellamy, Garland;Scott, C. Ronald
通讯作者：
Scott, C. Ronald
影响因子：
9.3
作者：
Li, YJ;Scott, CR;Gelb, MH
通讯作者：
Gelb, MH
影响因子：
9.3
作者：
De Jesus, Victor R.;Zhang, X. Kate;Hannon, W. Harry
通讯作者：
Hannon, W. Harry