Semiautomated high-sensitivity profiling of human blood serum glycoproteins through lectin preconcentration and multidimensional chromatography/tandem mass spectrometry

Milan Madera; Yehia Mechref; Iveta Klouckova; Milos V. Novotny

doi:10.1021/pr060169x

Semiautomated high-sensitivity profiling of human blood serum glycoproteins through lectin preconcentration and multidimensional chromatography/tandem mass spectrometry

Milan Madera, Yehia Mechref, Iveta Klouckova, Milos V. Novotny

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

69 Scopus citations

Abstract

We describe an effective analytical approach to identify trace glycoproteins in a small volume of human serum. The system is based on automatable affinity enrichment through silica-based lectin microcolumns and a further separation of the retained glycoproteins on a reversed-phase liquid chromatography with superficially porous packing, operating at high temperature. The fractionated sample is further directed into a 96-well plate for trypsinization and LC-MS/MS analysis. Using a major-component depleted blood serum (16 μg total protein), we were able to identify 271 glycoproteins through this analytical system.

Original language	English
Pages (from-to)	2348-2363
Number of pages	16
Journal	Journal of Proteome Research
Volume	5
Issue number	9
DOIs	https://doi.org/10.1021/pr060169x
State	Published - Sep 2006

Keywords

Glycoprotein Enrichment
Human Blood Serum Glycoproteins
LC-MS/MS
Microcolumn Lectin Affinity Chromatography
On-line Coupling of Microcolumn Lectin Affinity Chromatography and Poroshell LC-Columns
Protein Separation

Access to Document

10.1021/pr060169x

Cite this

@article{8f15ac8323b144499bad5f0af8ab4758,

title = "Semiautomated high-sensitivity profiling of human blood serum glycoproteins through lectin preconcentration and multidimensional chromatography/tandem mass spectrometry",

abstract = "We describe an effective analytical approach to identify trace glycoproteins in a small volume of human serum. The system is based on automatable affinity enrichment through silica-based lectin microcolumns and a further separation of the retained glycoproteins on a reversed-phase liquid chromatography with superficially porous packing, operating at high temperature. The fractionated sample is further directed into a 96-well plate for trypsinization and LC-MS/MS analysis. Using a major-component depleted blood serum (16 μg total protein), we were able to identify 271 glycoproteins through this analytical system.",

keywords = "Glycoprotein Enrichment, Human Blood Serum Glycoproteins, LC-MS/MS, Microcolumn Lectin Affinity Chromatography, On-line Coupling of Microcolumn Lectin Affinity Chromatography and Poroshell LC-Columns, Protein Separation",

author = "Milan Madera and Yehia Mechref and Iveta Klouckova and Novotny, {Milos V.}",

year = "2006",

month = sep,

doi = "10.1021/pr060169x",

language = "English",

volume = "5",

pages = "2348--2363",

journal = "Journal of Proteome Research",

issn = "1535-3893",

number = "9",

}

TY - JOUR

T1 - Semiautomated high-sensitivity profiling of human blood serum glycoproteins through lectin preconcentration and multidimensional chromatography/tandem mass spectrometry

AU - Madera, Milan

AU - Mechref, Yehia

AU - Klouckova, Iveta

AU - Novotny, Milos V.

PY - 2006/9

Y1 - 2006/9

N2 - We describe an effective analytical approach to identify trace glycoproteins in a small volume of human serum. The system is based on automatable affinity enrichment through silica-based lectin microcolumns and a further separation of the retained glycoproteins on a reversed-phase liquid chromatography with superficially porous packing, operating at high temperature. The fractionated sample is further directed into a 96-well plate for trypsinization and LC-MS/MS analysis. Using a major-component depleted blood serum (16 μg total protein), we were able to identify 271 glycoproteins through this analytical system.

AB - We describe an effective analytical approach to identify trace glycoproteins in a small volume of human serum. The system is based on automatable affinity enrichment through silica-based lectin microcolumns and a further separation of the retained glycoproteins on a reversed-phase liquid chromatography with superficially porous packing, operating at high temperature. The fractionated sample is further directed into a 96-well plate for trypsinization and LC-MS/MS analysis. Using a major-component depleted blood serum (16 μg total protein), we were able to identify 271 glycoproteins through this analytical system.

KW - Glycoprotein Enrichment

KW - Human Blood Serum Glycoproteins

KW - LC-MS/MS

KW - Microcolumn Lectin Affinity Chromatography

KW - On-line Coupling of Microcolumn Lectin Affinity Chromatography and Poroshell LC-Columns

KW - Protein Separation

UR - http://www.scopus.com/inward/record.url?scp=33748294165&partnerID=8YFLogxK

U2 - 10.1021/pr060169x

DO - 10.1021/pr060169x

M3 - Article

C2 - 16944947

AN - SCOPUS:33748294165

SN - 1535-3893

VL - 5

SP - 2348

EP - 2363

JO - Journal of Proteome Research

JF - Journal of Proteome Research

IS - 9

ER -

Semiautomated high-sensitivity profiling of human blood serum glycoproteins through lectin preconcentration and multidimensional chromatography/tandem mass spectrometry

Abstract

Keywords

Access to Document

Other files and links

Fingerprint

Cite this