Head-to-Head Comparison of Serum Fractionation Techniques

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• 作者：Jeffrey R. Whiteaker ; Heidi Zhang ; Jimmy K. Eng ; Ruihua Fang ; Brian D. Piening ; Li-Chia Feng ; Travis D. Lorentzen ; Regine M. Schoenherr ; John F. Keane ; Ted Holzman ; Matthew Fitzgibbon ; ChenWei Lin ; Hu
• 刊名：Journal of Proteome Research
• 出版年：2007
• 出版时间：February 2007
• 年：2007
• 卷：6
• 期：2
• 页码：828 - 836
• 全文大小：256K
• 年卷期：v.6,no.2(February 2007)
• ISSN：1535-3907

文摘

Multiple approaches for simplifying the serum proteome have been described. These techniques aregenerally developed across different laboratories, samples, mass spectrometry platforms, and analysistools. Hence, comparing the available schemes is impossible from the existing literature because ofconfounding variables. We describe a head-to-head comparison of several serum fractionation schemes,including N-linked glycopeptide enrichment, cysteinyl-peptide enrichment, magnetic bead separation(C3, C8, and WCX), size fractionation, protein A/G depletion, and immunoaffinity column depletion ofabundant serum proteins. Each technique was compared to results obtained from unfractionated humanserum. The results show immunoaffinity subtraction is the most effective means for simplifying theserum proteome while maintaining reasonable sample throughput. The reported dataset is publiclyavailable and provides a standard against which emergent technologies can be compared and evaluatedfor their contribution to serum-based biomarker discovery.