Analysis of the bovine plasma proteome by matrix-assisted laser desorption/ionisation time-of-flight tandem mass spectrometry

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• 作者：Ann-Kristin Henning ; Martin H. Groschup ; Thomas C. Mettenleiter ; Axel Karger
• 关键词：Bovine plasma ; Combinatorial peptide ligand library ; Proteomics ; MALDI-TOF/TOF mass spectrometry
• 刊名：The Veterinary Journal
• 出版年：January, 2014
• 年：2014
• 卷：199
• 期：1
• 页码：175-180
• 全文大小：428 K

文摘

In this study, the bovine plasma proteome was analysed using a three step protocol: (1) plasma was treated with a combinatorial peptide ligand library (CPLL) to assimilate the differences in concentrations of different proteins in raw plasma; (2) CPLL-treated material was fractionated by three standard electrophoretic separation techniques, and (3) samples were analysed by nano-liquid chromatography (nLC) matrix-assisted laser desorption/ionisation (MALDI) time-of-flight tandem (TOF/TOF) mass spectrometry. The efficiencies of three fractionation protocols for plasma proteome analysis were compared.

After size fractionation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), resolution of proteins was better and yields of identified proteins were higher than after charge-based fractionation by preparative gel-free isoelectric focussing. For proteins with isoelectric points >6 and molecular weights 猢?3 kDa, the best results were obtained with a 鈥榮hotgun鈥?approach, in which the CPLL-treated plasma was digested and the peptides, rather than the proteins, were fractionated by gel-free isoelectric focussing. However, the three fractionation techniques were largely complementary, since only about one-third of the proteome was identified by each approach.