Peptide mapping of proteins in cerebrospinal fluid utilizing a rapid preparative two-dimensional electrophoretic procedure and matrix-assisted laser desorption/ionization mass spectrometry

Pia Davidsson; Carol L. Nilsson

doi:10.1016/S0304-4165(99)00197-X

Peptide mapping of proteins in cerebrospinal fluid utilizing a rapid preparative two-dimensional electrophoretic procedure and matrix-assisted laser desorption/ionization mass spectrometry

Pia Davidsson, Carol L. Nilsson

Research output: Contribution to journal › Article › peer-review

37 Scopus citations

Abstract

A quick two-step procedure involving liquid phase isoelectric focusing in the Rotofor cell in combination with electroelution in the Mini whole cell gel eluter has been used for purification of proteins from human cerebrospinal fluid (CSF). Fractions, each highly enriched in a single protein band and virtually free of other proteins, were selected for characterization by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOFMS). Six CSF proteins, transferrin, α1-acid-glycoprotein, Zn-α2-glycoprotein, apolipoprotein A1, apolipoprotein E and β-trace were identified by MALDI-TOFMS analysis of the tryptic digests. These results demonstrate that the combination of liquid phase IEF and electroelution is a rapid preparative two-dimensional separation which can provide single proteins of high purity, in yields sufficient for characterization by MALDI-TOFMS. Characterization of such brain-specific proteins in CSF will be useful in the investigation of the pathophysiology of different brain disorders. Copyright (C) 1999 Elsevier Science B.V.

Original language	English (US)
Pages (from-to)	391-399
Number of pages	9
Journal	Biochimica et Biophysica Acta - General Subjects
Volume	1473
Issue number	2-3
DOIs	https://doi.org/10.1016/S0304-4165(99)00197-X
State	Published - Dec 27 1999
Externally published	Yes

Keywords

Cerebrospinal fluid
Electroelution
Liquid phase IEF
Matrix-assisted laser desorption/ionization mass spectrometry
Peptide mapping
Preparative two-dimensional gel electrophoresis

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology

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10.1016/S0304-4165(99)00197-X

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Peptide mapping of proteins in cerebrospinal fluid utilizing a rapid preparative two-dimensional electrophoretic procedure and matrix-assisted laser desorption/ionization mass spectrometry. / Davidsson, Pia; Nilsson, Carol L.
In: Biochimica et Biophysica Acta - General Subjects, Vol. 1473, No. 2-3, 27.12.1999, p. 391-399.

Research output: Contribution to journal › Article › peer-review

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title = "Peptide mapping of proteins in cerebrospinal fluid utilizing a rapid preparative two-dimensional electrophoretic procedure and matrix-assisted laser desorption/ionization mass spectrometry",

abstract = "A quick two-step procedure involving liquid phase isoelectric focusing in the Rotofor cell in combination with electroelution in the Mini whole cell gel eluter has been used for purification of proteins from human cerebrospinal fluid (CSF). Fractions, each highly enriched in a single protein band and virtually free of other proteins, were selected for characterization by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOFMS). Six CSF proteins, transferrin, α1-acid-glycoprotein, Zn-α2-glycoprotein, apolipoprotein A1, apolipoprotein E and β-trace were identified by MALDI-TOFMS analysis of the tryptic digests. These results demonstrate that the combination of liquid phase IEF and electroelution is a rapid preparative two-dimensional separation which can provide single proteins of high purity, in yields sufficient for characterization by MALDI-TOFMS. Characterization of such brain-specific proteins in CSF will be useful in the investigation of the pathophysiology of different brain disorders. Copyright (C) 1999 Elsevier Science B.V.",

keywords = "Cerebrospinal fluid, Electroelution, Liquid phase IEF, Matrix-assisted laser desorption/ionization mass spectrometry, Peptide mapping, Preparative two-dimensional gel electrophoresis",

author = "Pia Davidsson and Nilsson, {Carol L.}",

note = "Funding Information: This work was supported by grants from The Swedish Medical Research Council (Grants 12769, 13121); Foundation for Strategic Research (Infection and Vaccinology), Alzheimerfonden, Lund, Sweden; Bohuslandstingets FoU fond, Sweden; Stiftelsen Gamla Tj{\"a}nnarinnor, Stochholm, Sweden, Swedish Medical Society, Stiftelsen Handlanden Hjalmar Svenssons Forskningsfond, G{\"o}teborg, Sweden; Loo and Hans Osterman fond, Stockholm, Sweden; Lundbecksstiftelsen, Lund, Sweden; Wilhelm och Martina Lundgrens Stiftelse, G{\"o}teborg, Sweden, Magnus Bergvalls Stiftelse, Stockholm, Sweden; Thurings Stiftelse, Stockholm, Sweden, {\AA}ke Wibergs Stiftelse, Stockholm, Sweden. The authors would like to thank Thomas Larsson for participating in helpful discussions.",

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AU - Davidsson, Pia

AU - Nilsson, Carol L.

N1 - Funding Information: This work was supported by grants from The Swedish Medical Research Council (Grants 12769, 13121); Foundation for Strategic Research (Infection and Vaccinology), Alzheimerfonden, Lund, Sweden; Bohuslandstingets FoU fond, Sweden; Stiftelsen Gamla Tjännarinnor, Stochholm, Sweden, Swedish Medical Society, Stiftelsen Handlanden Hjalmar Svenssons Forskningsfond, Göteborg, Sweden; Loo and Hans Osterman fond, Stockholm, Sweden; Lundbecksstiftelsen, Lund, Sweden; Wilhelm och Martina Lundgrens Stiftelse, Göteborg, Sweden, Magnus Bergvalls Stiftelse, Stockholm, Sweden; Thurings Stiftelse, Stockholm, Sweden, Åke Wibergs Stiftelse, Stockholm, Sweden. The authors would like to thank Thomas Larsson for participating in helpful discussions.

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N2 - A quick two-step procedure involving liquid phase isoelectric focusing in the Rotofor cell in combination with electroelution in the Mini whole cell gel eluter has been used for purification of proteins from human cerebrospinal fluid (CSF). Fractions, each highly enriched in a single protein band and virtually free of other proteins, were selected for characterization by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOFMS). Six CSF proteins, transferrin, α1-acid-glycoprotein, Zn-α2-glycoprotein, apolipoprotein A1, apolipoprotein E and β-trace were identified by MALDI-TOFMS analysis of the tryptic digests. These results demonstrate that the combination of liquid phase IEF and electroelution is a rapid preparative two-dimensional separation which can provide single proteins of high purity, in yields sufficient for characterization by MALDI-TOFMS. Characterization of such brain-specific proteins in CSF will be useful in the investigation of the pathophysiology of different brain disorders. Copyright (C) 1999 Elsevier Science B.V.

AB - A quick two-step procedure involving liquid phase isoelectric focusing in the Rotofor cell in combination with electroelution in the Mini whole cell gel eluter has been used for purification of proteins from human cerebrospinal fluid (CSF). Fractions, each highly enriched in a single protein band and virtually free of other proteins, were selected for characterization by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOFMS). Six CSF proteins, transferrin, α1-acid-glycoprotein, Zn-α2-glycoprotein, apolipoprotein A1, apolipoprotein E and β-trace were identified by MALDI-TOFMS analysis of the tryptic digests. These results demonstrate that the combination of liquid phase IEF and electroelution is a rapid preparative two-dimensional separation which can provide single proteins of high purity, in yields sufficient for characterization by MALDI-TOFMS. Characterization of such brain-specific proteins in CSF will be useful in the investigation of the pathophysiology of different brain disorders. Copyright (C) 1999 Elsevier Science B.V.

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Peptide mapping of proteins in cerebrospinal fluid utilizing a rapid preparative two-dimensional electrophoretic procedure and matrix-assisted laser desorption/ionization mass spectrometry

Abstract

Keywords

ASJC Scopus subject areas

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