An Integrated Workflow for Multiplex CSF Proteomics and Peptidomics鈥擨dentification of Candidate Cerebrospinal Fluid Biomarkers of Alzheimer鈥檚 Disease

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• 作者：Mikko H枚ltt盲 ; Lennart Minthon ; Oskar Hansson ; Jessica Holm茅n-Larsson ; Ian Pike ; Malcolm Ward ; Karsten Kuhn ; Ulla R眉etschi ; Henrik Zetterberg ; Kaj Blennow ; Johan Gobom
• 刊名：Journal of Proteome Research
• 出版年：2015
• 出版时间：February 6, 2015
• 年：2015
• 卷：14
• 期：2
• 页码：654-663
• 全文大小：274K
• ISSN：1535-3907

文摘

Many disease processes in the brain are reflected in the protein composition of the cerebrospinal fluid (CSF). In addition to proteins, CSF also contains a large number of endogenous peptides whose potential as disease biomarkers largely remains to be explored. We have developed a novel workflow in which multiplex isobaric labeling is used for simultaneous quantification of endogenous CSF peptides and proteins by liquid chromatography coupled with mass spectrometry. After the labeling of CSF samples, endogenous peptides are separated from proteins by ultrafiltration. The proteins retained on the filters are trypsinized, and the tryptic peptides are collected separately. We evaluated this technique in a comparative pilot study of CSF peptide and protein profiles in eight patients with Alzheimer鈥檚 disease (AD) and eight nondemented controls. We identified several differences between the AD and control group among endogenous peptides derived from proteins known to be associated with AD, including neurosecretory protein VGF (ratios AD/controls 0.45鈥?.81), integral membrane protein 2B (ratios AD/controls 0.72鈥?.84), and metallothionein-3 (ratios AD/controls 0.51鈥?.61). Analysis of tryptic peptides identified several proteins that were altered in the AD group, some of which have previously been reported as changed in AD, for example, VGF (ratio AD/controls 0.70).