Improved resolution of human cerebrospinal fluid proteins on two-dimensional gels.
Proteomics combines two-dimensional gel electrophoresis and peptide mass fingerprinting and can potentially identify a protein(s) unique to disease. Such proteins can be used either for diagnosis or may be relevant to the pathogenesis of disease. Because patients with multiple sclerosis (MS) have increased amounts of immunoglobulin (Ig) G in their cerebrospinal fluid (CSF) that is directed against an as yet unidentified protein, we are applying proteomics to MS CSF, studies that require optimal separation of proteins in human CSF. We found that recovery of proteins from CSF of MS patients was improved using ultrafiltration, rather than dialysis, for desalting. Resolution of these proteins was enhanced by acetone precipitation of desalted CSF before electrophoresis and by fractionation of CSF using Cibacron Blue sepharose affinity chromatography. Improved protein recovery and resolution will facilitate excision from gels for analysis by peptide mass fingerprinting.
Department of Neurology, University of Colorado Health Sciences Center, Denver, Colorado 80262, USA. barbara.hammack@uchsc.edu
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Hammack
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Owens
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Burgoon
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D000418:Albumins; D002556:Cerebrospinal Fluid Proteins / Q000032:analysis*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods; D009103:Multiple Sclerosis / Q000134:cerebrospinal fluid*; D012492:Salts
D000418:Albumins; D002556:Cerebrospinal Fluid Proteins; D012492:Salts