Standardized approach to proteome profiling of human serum based on magnetic bead separation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

被引：188

作者：

Baumann, S ^{[1
]}

Ceglarek, U ^{[1
]}

Fiedler, GM ^{[1
]}

Lembcke, J ^{[1
]}

Leichtle, A ^{[1
]}

Thiery, J ^{[1
]}

机构：

[1] Univ Hosp Leipzig, Inst Lab Med Clin Chem & Mol Diagnost, D-04103 Leipzig, Germany

来源：

CLINICAL CHEMISTRY | 2005年 / 51卷 / 06期

关键词：

D O I：

10.1373/clinchem.2004.047308

中图分类号：

R446 [实验室诊断]; R-33 [实验医学、医学实验];

学科分类号：

1001 ;

摘要：

Background: Magnetic bead purification for the analysis of low-abundance proteins in body fluids facilitates the identification of potential new biomarkers by matrix-assisted laser des orption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The aims of our study were to establish a proteome fractionation technique and to validate a standardized blood sampling, processing, and storage procedure for proteomic pattern analysis. Methods: We used magnetic bead separation for proteome profiling of human blood by MALDI-TOF MS (mass range, 1000-10000 Da) and studied the effects on the quality and reproducibility of the proteome analysis of anticoagulants, blood clotting, time and temperature of sample storage, and the number of freeze-thaw cycles of samples. Results: The proteome pattern of human serum was characterized by similar to 350 signals in the mass range of 1000-10000 Da. The proteome profile showed time-dependent dynamic changes before and after centrifugation of the blood samples. Serum mass patterns differed between native samples and samples frozen once. The best reproducibility of proteomic patterns was with a single thawing of frozen serum samples. Conclusion: Application of the standardized preanalytical blood sampling and storage procedure in combination with magnetic bead-based fractionation decreases variability of proteome patterns in human serum assessed by MALDI-TOF MS. (c) 2005 American Association for Clinical Chemistry

引用

页码：973 / 980

页数：8

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