Improving limits of detection for B-type natriuretic peptide using PC-IDMS: an application of the ALiPHAT strategy.
Improving limits of detection for B-type natriuretic peptide using PC-IDMS: an application of the ALiPHAT strategy.
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DOI：
10.1039/b919484c
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发表时间：
2010-01
期刊：
影响因子：
--
通讯作者：
Muddiman DC
中科院分区：
文献类型：
--
作者：
Shuford CM;Comins DL;Whitten JL;Burnett JC Jr;Muddiman DC
我们的小组和其他人已经报道了生物分子的疏水标记，以增加它们在电喷雾电离过程中的电离效率，并促进它们通过质谱法的检测。因此，疏水标记应该提供一种可行的方法，用于通过降低其检测限来增强低丰度蛋白质的基于MS的定量。在此，我们评估了两种商业烷基化试剂和几种新合成的疏水烷基化试剂，用于通过蛋白裂解同位素稀释质谱法定量B型利钠肽（一种低丰度心脏生物标志物）。对于评价的含半胱氨酸的胰蛋白酶肽，相对于常用的烷基化试剂碘乙酰胺，观察到性能最佳的疏水试剂2-碘-N-辛基乙酰胺的检测限降低约3.5倍。此外，我们还评估了使用非极性表面积作为衡量烷基化试剂在改善ESI响应方面的有效性的指标。
Hydrophobic tagging of biomolecules has been reported by our group and others to increase their ionization efficiency during electrospray ionization and facilitate their detection by mass spectrometry. As such, hydrophobic tagging should provide a viable method for augmenting MS-based quantification of low abundance proteins by decreasing their detection limits. Herein we have evaluated two commercial alkylation reagents and several newly synthesized hydrophobic alkylation reagents for their utility in quantifying B-type Natriuretic Peptide, a low abundance cardiac biomarker, by protein cleavage isotope dilution mass spectrometry. For the cysteine containing tryptic peptide evaluated, a ∼3.5-fold decrease in the detection limit was observed for the best performing hydrophobic reagent, 2-iodo-N-octylacetamide, relative to the commonly used alkylation reagent, iodoacetamide. Additionally, we have evaluated the use of nonpolar surface areas as a metric for assessing the effectiveness of the alkylation reagents in improving ESI response.
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DOI：
10.1016/j.jchromb.2009.02.040
发表时间：
2009-04-01
影响因子：
3
作者：
Andrews, Genna L.;Shuford, Christopher M.;Burnett, John C., Jr.;Hawkridge, Adam M.;Muddiman, David C.
通讯作者：
Muddiman, David C.
影响因子：
4.4
作者：
Anderson, NL;Anderson, NG;Pearson, TW
通讯作者：
Pearson, TW
影响因子：
158.5
作者：
Maisel, AS;Krishnaswamy, P;McCullough, PA
通讯作者：
McCullough, PA
影响因子：
7.4
作者：
Mirzaei, Hamid;Regnier, Fred
通讯作者：
Regnier, Fred
DOI：
10.1073/pnas.0508782102
发表时间：
2005-11-29
影响因子：
11.1
作者：
Hawkridge, AM;Heublein, DM;Muddiman, DC
通讯作者：
Muddiman, DC