Feasibility study using surface-enhanced Raman spectroscopy for the quantitative detection of tyrosine and serine phosphorylation
Feasibility study using surface-enhanced Raman spectroscopy for the quantitative detection of tyrosine and serine phosphorylation
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DOI：
10.1016/j.bbagen.2007.01.018
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发表时间：
2007-06-01
影响因子：
3
通讯作者：
Winlove, C. P.
中科院分区：
文献类型：
--
作者：
Moger, J.;Gribbon, P.;Winlove, C. P.
我们研究了基于胶体的表面增强拉曼散射（Sers）作为一种高灵敏度的技术检测丝氨酸和酪氨酸残基的肽磷酸化的可行性。使用最近报道的滴涂沉积拉曼方法，我们验证我们的Sers光谱对正常的拉曼光谱，否则将无法获得在这样低的浓度。与现有的肽磷酸化定量技术（如高效液相色谱法（HPLC））相比，与Sers相关的短扫描和处理时间使其成为亚微摩尔浓度近实时测量的有吸引力的替代方案。在通过Savistky-Golay二阶导数（SGSD）预处理之后，使用多变量光谱分类、区间偏最小二乘法（iPLS）确定合成肽的磷酸化程度。此外，我们的研究结果表明，该技术是强大的干扰，从复杂的蛋白质和其他磷酸化的化合物存在的浓度通常发现在筛选试验。(c)2007 Elsevier B. V.保留所有权利。
We investigate the feasibility of colloid-based surface enhanced Raman scattering (SERS) as a highly sensitive technique for detecting peptide phosphorylation at serine and tyrosine residues. Using the recently reported drop-coating deposition Raman method we validate our SERS spectra against normal Raman spectra that would otherwise be unobtainable at such low concentrations. Compared with existing techniques for quantifying peptide phosphorylation, such as high-performance liquid chromatography (HPLC), the short scanning and processing time associated with SERS makes it an attractive alternative for near-real-time measurement at sub micro-molar concentrations. Following pre-processing by Savistky-Golay second derivative (SGSD), the degree of phosphorylation of synthetic peptides is determined using multivariate spectral classification, interval partial least squares (iPLS). Furthermore, our results show that the technique is robust to interference from complex proteins and other phosphorylated compounds present at concentrations typically found in a screening assay. (c) 2007 Elsevier B.V. All rights reserved.