MALDI-target integrated platform for affinity-captured protein digestion

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• 作者：Asilah Ahmad-Tajudin ; Belinda Adler ; Simon Ekstr枚m ; Gy枚rgy Marko-Varga ; Johan Malm ; Hans Lilja ; Thomas Laurell
• 关键词：MS ; mass spectrometry ; MRM ; multiple reaction monitoring ; MALDI ; matrix-assisted laser desorption/ionization ; SPR ; surface plasmon resonance ; ISET ; integrated selective enrichment target ; SPE ; solid-phase extraction ; RP ; reverse phase ; PSA ; prostate specific antigen
• 刊名：Analytica Chimica Acta
• 出版年：7 January, 2014
• 年：2014
• 卷：807
• 期：Complete
• 页码：1-8
• 全文大小：1987 K

文摘

To address immunocapture of proteins in large cohorts of clinical samples high throughput sample processing is required. Here a method using the proteomic sample platform, ISET (integrated selective enrichment target) that integrates highly specific immunoaffinity capture of protein biomarker, digestion and sample cleanup with a direct interface to mass spectrometry is presented. The robustness of the on-ISET protein digestion protocol was validated by MALDI MS analysis of model proteins, ranging from 40 fmol to 1 pmol per nanovial. On-ISET digestion and MALDI MS/MS analysis of immunoaffinity captured disease-associated biomarker PSA (prostate specific antigen) from human seminal plasma are presented.