Applications of monolithic silica capillary columns in proteomics

被引:42
作者
Barroso, B
Lubda, D
Bischoff, R
机构
[1] Univ Groningen, Dept Bioanal & Toxicol, Univ Ctr Pharm, NL-9713 AV Groningen, Netherlands
[2] Merck KGaA, LSP R&D MDA, D-64271 Darmstadt, Germany
关键词
proteomics; monoliths; Bronchoalveolar Lavage Fluid; serum; cancer; mass spectrometry; HPLC;
D O I
10.1021/pr0340532
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The use and applicability of silica based capillary monolithic reversed-phase columns in proteomic analysis has been evaluated by liquid chromatography-mass spectrometry (LC-MS). Chromatographic performance of the monolithic capillaries was evaluated with a tryptic digest of cytochrome C showing very good resolution and reproducibility in addition to the known advantages of a low pressure drop over a time period of 6 months. Monoliths were subsequently tested for their suitability to separate proteins and peptides from samples typically encountered in proteomic research such as in-gel digested tryptic peptide mixtures or fractions of proteolytically digested human serum. The monolithic capillaries also proved useful in the analysis of phospholipid species in bronchoalveolar lavage fluid. Compared to particle-filled conventional capillary columns, rapid and highly efficient separation of peptides and proteins was achieved using these bimodal pore size distribution columns, and good quality collision induced dissociation (CID) mass spectra were obtained on an ion trap mass spectrometer. These novel monolithic separation media are thus a promising addition to the methodological toolbox of proteomics research.
引用
收藏
页码:633 / 642
页数:10
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