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Molecular & Cellular Proteomics 6:755-766, 2007.
© 2007 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

Analysis and Quantification of Diagnostic Serum Markers and Protein Signatures for Gaucher Disease^*,S

Johannes P. C. Vissers^,, James I. Langridge and Johannes M. F. G. Aerts^¶

From the Waters Corporation, MS Technologies Center, Manchester M22 5PP, United Kingdom and ^¶ Department of Biochemistry, Academic Medical Center, University of Amsterdam, Amsterdam, 1105 AZ, The Netherlands

Novel approaches for the qualitative and quantitative proteomics analysis by nanoscale LC-MS applied to the study of protein expression response in depleted and undepleted serum of Gaucher patients undergoing enzyme replacement therapy are presented. Particular emphasis is given to the method reproducibility of these LC-MS experiments without the use of isotopic labels. The level of chitotriosidase, an established Gaucher biomarker, was assessed by means of an absolute concentration determination technique for alternate scanning LC-MS generated data. Disease associated proteins, including fibrinogens, complement cascade proteins, and members of the high density lipoprotein serum content, were recognized by various clustering methods and sorting and intensity profile grouping of identified peptides. Condition-unique LC-MS protein signatures could be generated utilizing the measured serum protein concentrations and are presented for all investigated conditions. The clustering results of the study were also used as input for gene ontology searches to determine the correlation between the molecular functions of the identified peptides and proteins.

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