Multidimensional protein fractionation using ProteomeLab PF 2D™ for profiling amyotrophic lateral sclerosis immunity: A preliminary reportReportar como inadecuado




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Proteome Science

, 6:26

First Online: 12 September 2008Received: 01 April 2008Accepted: 12 September 2008

Abstract

BackgroundThe ProteomeLab™ PF 2D platform is a relatively new approach to global protein profiling. Herein, it was used for investigation of plasma proteome changes in amyotrophic lateral sclerosis ALS patients before and during immunization with glatiramer acetate GA in a clinical trial.

ResultsThe experimental design included immunoaffinity depletion of 12 most abundant proteins from plasma samples with the ProteomeLab™ IgY-12 LC10 column kit as first dimension separation, also referred to as immuno-partitioning. Second and third dimension separations of the enriched proteome were performed on the PF 2D platform utilizing 2D isoelectric focusing and RP-HPLC with the resulting fractions collected for analysis. 1D gel electrophoresis was added as a fourth dimension when sufficient protein was available. Protein identification from collected fractions was performed using nano-LC-MS-MS approach. Analysis of differences in the resulting two-dimensional maps of fractions obtained from the PF 2D and the ability to identify proteins from these fractions allowed sensitivity threshold measurements. Masked proteins in the PF 2D fractions are discussed.

ConclusionWe offer some insight into the strengths and limitations of this emerging proteomic platform.

Electronic supplementary materialThe online version of this article doi:10.1186-1477-5956-6-26 contains supplementary material, which is available to authorized users.

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Autor: Joshua D Schlautman - Wojciech Rozek - Robert Stetler - R Lee Mosley - Howard E Gendelman - Pawel Ciborowski

Fuente: https://link.springer.com/







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