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Microwave and magnetic (M 2 ) proteomics of the experimental autoimmune encephalomyelitis animal model of multiple sclerosis
Author(s) -
Raphael Itay,
Mahesula Swetha,
Kalsaria Karan,
Kotagiri Venkat,
Purkar Anjali B.,
Anjanappa Manjushree,
Shah Darshit,
Pericherla Vidya,
Jadhav Yeshwant Lal Avinash,
Raghunathan Rekha,
Vaynberg Michael,
Noriega David,
Grimaldo Nazul H.,
Wenk Carola,
Gelfond Jonathan A.L.,
Forsthuber Thomas G.,
Haskins William E.
Publication year - 2012
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201200200
Subject(s) - multiple sclerosis , experimental autoimmune encephalomyelitis , encephalomyelitis , proteomics , nuclear magnetic resonance , microwave , animal model , chemistry , physics , medicine , immunology , biochemistry , gene , quantum mechanics
We hypothesized that quantitative MS / MS ‐based proteomics at multiple time points, incorporating rapid microwave and magnetic ( M 2 ) sample preparation, could enable relative protein expression to be correlated to disease progression in the experimental autoimmune encephalomyelitis ( EAE ) animal model of multiple sclerosis. To test our hypothesis, microwave‐assisted reduction/alkylation/digestion of proteins from brain tissue lysates bound to C 8 magnetic beads and microwave‐assisted isobaric chemical labeling were performed of released peptides, in 90 s prior to unbiased proteomic analysis. Disease progression in EAE was assessed by scoring clinical EAE disease severity and confirmed by histopathologic evaluation for central nervous system inflammation. Decoding the expression of 283 top‐ranked proteins ( p <0.05) at each time point relative to their expression at the peak of disease, from a total of 1191 proteins observed in four technical replicates, revealed a strong statistical correlation to EAE disease score, particularly for the following four proteins that closely mirror disease progression: 14‐3‐3ε ( p = 3.4 E ‐6); GPI ( p = 2.1 E ‐5); PLP 1 ( p = 8.0 E ‐4); PRX 1 ( p = 1.7 E ‐4). These results were confirmed by W estern blotting, signaling pathway analysis, and hierarchical clustering of EAE risk groups. While validation in a larger cohort is underway, we conclude that M 2 proteomics is a rapid method to quantify putative prognostic/predictive protein biomarkers and therapeutic targets of disease progression in the EAE animal model of multiple sclerosis.