Head-to-Head Comparison of Serum Fractionation Techniques
Autor: | Jimmy K. Eng, Leigh Anderson, Heidi Zhang, Ted Holzman, Li Chia Feng, Tao Liu, Richard D. Smith, Martin W. McIntosh, Brian D. Piening, J.F. Keane, Regine M. Schoenherr, Chen Wei Lin, Amanda G. Paulovich, Kelly Cooke, Jeffrey R. Whiteaker, Julian D. Watts, Travis D. Lorentzen, Matthew Fitzgibbon, David G. Camp, Ruihua Fang, Hui Zhang |
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Rok vydání: | 2006 |
Předmět: |
Male
Proteomics Spectrometry Mass Electrospray Ionization Chromatography biology Chemistry Head to head Glycopeptides Blood Proteins General Chemistry Fractionation Mass spectrometry Biochemistry Blood proteins Mass Spectrometry Peptide Fragments Magnetic Bead Separation Liquid chromatography–mass spectrometry biology.protein Humans Trypsin Biomarker discovery Protein A Chromatography Liquid |
Zdroj: | Journal of Proteome Research. 6:828-836 |
ISSN: | 1535-3907 1535-3893 |
Popis: | Multiple approaches for simplifying the serum proteome have been described. These techniques are generally developed across different laboratories, samples, mass spectrometry platforms, and analysis tools. Hence, comparing the available schemes is impossible from the existing literature because of confounding 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 of abundant serum proteins. Each technique was compared to results obtained from unfractionated human serum. The results show immunoaffinity subtraction is the most effective means for simplifying the serum proteome while maintaining reasonable sample throughput. The reported dataset is publicly available and provides a standard against which emergent technologies can be compared and evaluated for their contribution to serum-based biomarker discovery. |
Databáze: | OpenAIRE |
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