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
Rok vydání: 2006
Předmět:
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