基于聚（甲基丙烯酸缩水甘油酯-二乙烯基苯）的固定化 pH 梯度毛细管等电聚焦与 MALDI 质谱联用，用于增强神经肽分析。
Poly(glycidyl methacrylate-divinylbenzene) based immobilized pH gradient capillary isoelectric focusing coupling with MALDI mass spectrometry for enhanced neuropeptide analysis.
机构信息
School of Pharmacy, University of Wisconsin, Madison, WI, USA.
出版信息
Electrophoresis. 2012 Feb;33(4):661-5. doi: 10.1002/elps.201100447.
Abstract
Herein, we report an immobilized pH gradient (IPG) capillary isoelectric focusing-matrix-assisted laser desorption/ionization mass spectrometry (CIEF-MALDI MS) platform designed for the separation of complex neuropeptides. This platform features a poly(glycidyl methacrylate-divinylbenzene) (GMA-DVB)-based monolithic column for CIEF separation. Different from regular CIEF, carrier ampholytes are preimmobilized on the monolithic surface instead of being added to the sample. An off-line coupling of IPG-CIEF to MALDI MS has been established. Comparison with regular CIEF and optimizations are performed with bovine serum albumin tryptic peptides and extracted neuropeptide mixtures from crustacean Callinectes sapidus. It has been demonstrated that the separation of complex peptide mixtures in neutral and basic pH ranges can be achieved in less than 10 min with comparable separation efficiency with regular CIEF, while the MS signal is significantly enhanced when employing IPG-CIEF. Enhanced neuropeptide detection is also observed after coupling IPG-CIEF with MALDI MS.
摘要
本文报道了一种固定化 pH 梯度（IPG）毛细管等电聚焦-基质辅助激光解吸/电离质谱（CIEF-MALDI MS）平台，用于分离复杂的神经肽。该平台的特点是采用聚（甲基丙烯酸缩水甘油酯-二乙烯基苯）（GMA-DVB）为基质的整体柱进行 CIEF 分离。与常规 CIEF 不同，载体两性电解质预先固定在整体柱表面，而不是添加到样品中。建立了 IPG-CIEF 与 MALDI MS 的离线耦合。采用牛血清白蛋白胰蛋白酶肽和甲壳类动物中国对虾提取的神经肽混合物进行比较和优化。结果表明，在中性和碱性 pH 范围内，复杂肽混合物的分离可以在不到 10 分钟内完成，与常规 CIEF 相比具有相当的分离效率，而采用 IPG-CIEF 时，MS 信号显著增强。将 IPG-CIEF 与 MALDI MS 耦合后，还观察到神经肽的检测得到增强。
相似文献
1
Poly(glycidyl methacrylate-divinylbenzene) based immobilized pH gradient capillary isoelectric focusing coupling with MALDI mass spectrometry for enhanced neuropeptide analysis.基于聚（甲基丙烯酸缩水甘油酯-二乙烯基苯）的固定化 pH 梯度毛细管等电聚焦与 MALDI 质谱联用，用于增强神经肽分析。
Electrophoresis. 2012 Feb;33(4):661-5. doi: 10.1002/elps.201100447.
2
Membrane-assisted capillary isoelectric focusing coupling with matrix-assisted laser desorption/ionization-Fourier transform mass spectrometry for neuropeptide analysis.膜辅助毛细管等电聚焦与基质辅助激光解吸/电离-傅里叶变换质谱联用分析神经肽。
J Chromatogr A. 2011 Aug 5;1218(31):5336-43. doi: 10.1016/j.chroma.2011.05.072. Epub 2011 May 27.
3
Preparation of a capillary isoelectric focusing column with monolithic immobilized pH gradient and its application on protein separation based on an online capillary isoelectric focusing platform.制备具有整体固定化 pH 梯度的毛细管等电聚焦柱及其在基于在线毛细管等电聚焦平台的蛋白质分离中的应用。
Electrophoresis. 2019 Jul;40(12-13):1722-1730. doi: 10.1002/elps.201900125. Epub 2019 Apr 29.
4
Capillary isoelectric focusing of probiotic bacteria from cow's milk in tapered fused silica capillary with off-line matrix-assisted laser desorption/ionization time-of-flight mass spectrometry identification.牛乳制品中益生菌的毛细管等电聚焦与离线基质辅助激光解吸/电离飞行时间质谱鉴定。
Anal Chim Acta. 2013 Jul 25;788:193-9. doi: 10.1016/j.aca.2013.05.059. Epub 2013 Jun 10.
5
Capillary isoelectric focusing with free or immobilized pH gradient in silica particles packed column.毛细等电聚焦与自由或固定 pH 梯度在硅胶颗粒填充柱中。
Anal Chim Acta. 2019 Nov 4;1079:230-236. doi: 10.1016/j.aca.2019.06.013. Epub 2019 Jun 8.
6
Ultra-fast mass fingerprinting by high-affinity capture of peptides and proteins on derivatized poly(glycidyl methacrylate/divinylbenzene) for the analysis of serum and cell lysates.通过在衍生化聚（甲基丙烯酸缩水甘油酯/二乙烯基苯）上高亲和力捕获肽和蛋白质进行超快速质量指纹分析，用于血清和细胞裂解物分析。
Rapid Commun Mass Spectrom. 2006;20(19):2954-60. doi: 10.1002/rcm.2673.
7
On-line combination of monolithic immobilized pH gradient-based capillary isoelectric focusing and capillary zone electrophoresis via a partially etched porous interface for protein analysis.通过部分刻蚀多孔界面将基于整体固定化 pH 梯度的毛细管等电聚焦和毛细管区带电泳在线组合用于蛋白质分析。
J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Apr 1;879(11-12):804-10. doi: 10.1016/j.jchromb.2011.02.020. Epub 2011 Feb 21.
8
Capillary isoelectric focusing hyphenated to single- and multistage matrix-assisted laser desorption/ionization-mass spectrometry using automated sheath-flow-assisted sample deposition.毛细管等电聚焦联用单级和多级基质辅助激光解吸/电离质谱，采用自动鞘流辅助样品沉积。
Electrophoresis. 2008 May;29(10):1974-84. doi: 10.1002/elps.200700836.
9
Combination of capillary isoelectric focusing in a tapered capillary with MALDI-TOF MS for rapid and reliable identification of Dickeya species from plant samples.毛细管等电聚焦与 MALDI-TOF MS 的组合在锥形毛细管中用于快速可靠地鉴定来自植物样本的迪氏棒状杆菌属种。
Anal Chem. 2013 Jul 16;85(14):6806-12. doi: 10.1021/ac4009176. Epub 2013 Jul 2.
10
UV embossed polymeric chip for protein separation and identification based on capillary isoelectric focusing and MALDI-TOF-MS.基于毛细管等电聚焦和基质辅助激光解吸电离飞行时间质谱的用于蛋白质分离与鉴定的紫外压纹聚合物芯片
Anal Chem. 2006 May 15;78(10):3249-56. doi: 10.1021/ac051773e.
引用本文的文献
1
Recent advances in the analysis of therapeutic proteins by capillary and microchip electrophoresis.毛细管电泳和微芯片电泳在治疗性蛋白质分析中的最新进展。
Anal Methods. 2014 Jul 8;6(15):5427-5449. doi: 10.1039/C4AY00447G.
2
Recent advances in coupling capillary electrophoresis-based separation techniques to ESI and MALDI-MS.基于毛细管电泳的分离技术与电喷雾电离和基质辅助激光解吸电离质谱联用的最新进展。
Electrophoresis. 2014 May;35(9):1214-25. doi: 10.1002/elps.201300451. Epub 2013 Dec 5.
3
Small-volume analysis of cell-cell signaling molecules in the brain.大脑中细胞间信号分子的小体积分析
Neuropsychopharmacology. 2014 Jan;39(1):50-64. doi: 10.1038/npp.2013.145. Epub 2013 Jun 10.
4
Semi-automated liquid chromatography-mass spectrometric imaging platform for enhanced detection and improved data analysis of complex peptides.用于增强复杂肽检测和改善数据分析的半自动化液相色谱-质谱成像平台。
J Chromatogr A. 2013 Jun 7;1293:44-50. doi: 10.1016/j.chroma.2013.03.042. Epub 2013 Mar 26.
5
Trajectory of isoelectric focusing from gels to capillaries to immobilized gradients in capillaries.从凝胶到毛细管再到毛细管中固定梯度的等电聚焦轨迹。
Proteomics. 2012 Oct;12(19-20):2918-26. doi: 10.1002/pmic.201200213. Epub 2012 Sep 24.
6
Neuropeptide analysis with liquid chromatography-capillary electrophoresis-mass spectrometric imaging.采用液相色谱-毛细管电泳-质谱成像技术进行神经肽分析。
J Sep Sci. 2012 Jul;35(14):1779-84. doi: 10.1002/jssc.201200051.
本文引用的文献
1
Membrane-assisted capillary isoelectric focusing coupling with matrix-assisted laser desorption/ionization-Fourier transform mass spectrometry for neuropeptide analysis.膜辅助毛细管等电聚焦与基质辅助激光解吸/电离-傅里叶变换质谱联用分析神经肽。
J Chromatogr A. 2011 Aug 5;1218(31):5336-43. doi: 10.1016/j.chroma.2011.05.072. Epub 2011 May 27.
2
Advancing matrix-assisted laser desorption/ionization-mass spectrometric imaging for capillary electrophoresis analysis of peptides.推进基质辅助激光解吸/电离质谱成像在毛细管电泳分析肽中的应用。
Anal Chem. 2011 May 1;83(9):3462-9. doi: 10.1021/ac200708f. Epub 2011 Apr 6.
3
Analysis of low molecular weight acids by monolithic immobilized pH gradient-based capillary isoelectric focusing coupled with mass spectrometry.基于整体固定化 pH 梯度的毛细管等电聚焦与质谱联用分析低分子量酸。
J Sep Sci. 2011 Feb;34(4):422-7. doi: 10.1002/jssc.201000720. Epub 2011 Jan 24.
4
Integration of capillary isoelectric focusing with monolithic immobilized pH gradient, immobilized trypsin microreactor and capillary zone electrophoresis for on-line protein analysis.毛细管等电聚焦与整体固定化 pH 梯度、固定化胰蛋白酶微反应器和毛细管区带电泳的在线蛋白质分析集成。
J Sep Sci. 2010 Oct;33(20):3194-200. doi: 10.1002/jssc.201000324.
5
Peptides in the brain: mass spectrometry-based measurement approaches and challenges.脑内肽：基于质谱的测量方法及挑战。
Annu Rev Anal Chem (Palo Alto Calif). 2008;1:451-83. doi: 10.1146/annurev.anchem.1.031207.113053.
6
Recent progress of chiral monolithic stationary phases in CEC and capillary LC.手性整体固定相在 CEC 和毛细管 LC 中的最新进展。
Electrophoresis. 2010 May;31(9):1457-66. doi: 10.1002/elps.200900672.
7
Combining capillary electrophoresis matrix-assisted laser desorption/ionization mass spectrometry and stable isotopic labeling techniques for comparative crustacean peptidomics.结合毛细管电泳基质辅助激光解吸/电离质谱和稳定同位素标记技术进行甲壳动物比较肽组学研究。
J Chromatogr A. 2010 Jun 25;1217(26):4463-70. doi: 10.1016/j.chroma.2010.02.084. Epub 2010 Mar 6.
8
A systematic study in CIEF: defining and optimizing experimental parameters critical to method reproducibility and robustness.在 CIEF 中的系统研究：定义和优化对方法重现性和稳健性至关重要的实验参数。
Electrophoresis. 2009 Dec;30(23):4049-58. doi: 10.1002/elps.200800690.
9
Microchip isoelectric focusing with monolithic immobilized pH gradient materials for proteins separation.微芯片等电聚焦与整体固定化 pH 梯度材料在蛋白质分离中的应用。
Electrophoresis. 2009 Dec;30(23):4034-9. doi: 10.1002/elps.200900209.
10
Mass spectrometric characterization and physiological actions of novel crustacean C-type allatostatins.新型甲壳类C型咽侧体抑制素的质谱表征及生理作用
Peptides. 2009 Sep;30(9):1660-8. doi: 10.1016/j.peptides.2009.05.023. Epub 2009 Jun 6.
Zotero 插件