Microwave and magnetic (M2) proteomics of the experimental autoimmune encephalomyelitis animal model of multiple sclerosis

We hypothesized that quantitative MS/MS‐based proteomics at multiple time points, incorporating rapid microwave and magnetic (M2) sample preparation, could enable relative protein expression to be correlated to disease progression in the experimental autoimmune encephalomyelitis (EAE) animal model o...

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Published in:Electrophoresis 2012-12, Vol.33 (24), p.3810-3819
Main Authors: 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.
Format: Article
Language:English
Subjects:
Animals
Beads
Blotting, Western
Brain - metabolism
Brain - physiopathology
Brain Chemistry
Central nervous system
Cluster Analysis
Disease Models, Animal
Encephalomyelitis, Autoimmune, Experimental - metabolism
Female
Isobaric chemical labeling
Magnetics
Mice
Mice, Inbred C57BL
Microwave proteomics
Microwaves
Multiple sclerosis
Multiple Sclerosis - metabolism
Progressions
Proteins
Proteome - analysis
Proteome - metabolism
Proteomics
Proteomics - methods
Sample preparation
Tandem Mass Spectrometry - methods
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Summary:We hypothesized that quantitative MS/MS‐based proteomics at multiple time points, incorporating rapid microwave and magnetic (M2) 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 C8 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
ISSN:0173-0835
1522-2683
DOI:10.1002/elps.201200200