Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis

In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [12C]- and [13C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants i...

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Autores: Giménez, Estela, Balmaña, Meritxell, Figueras, Joan, Fort Martorell, Esther, Bolòs, Carme de, Sanz Nebot, María Victoria, Peracaula Miró, Rosa, Rizzi, Andreas
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2015
País:España
Institución:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:10256/12672
Acceso en línea:http://hdl.handle.net/10256/12672
Access Level:acceso embargado
Palabra clave:Pàncrees -- Càncer
Pancreas -- Cancer
Glicoproteïnes
Glycoproteins
Espectrometria de masses
Mass spectrometry
Marcadors tumorals
Tumor markers
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spelling Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosisGiménez, EstelaBalmaña, MeritxellFigueras, JoanFort Martorell, EstherBolòs, Carme deSanz Nebot, María VictoriaPeracaula Miró, RosaRizzi, AndreasPàncrees -- CàncerPancreas -- CancerGlicoproteïnesGlycoproteinsEspectrometria de massesMass spectrometryMarcadors tumoralsTumor markersIn this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [12C]- and [13C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. Human α1-acid-glycoprotein (hAGP) is an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation. However, it is not clear yet whether some particular glycans in hAGP can be used as biomarker for differentiating between these two pathologies. In this work, hAGP was isolated by immunoaffinity chromatography (IAC) from serum samples of healthy individuals and from those suffering chronic pancreatitis and different stages of pancreatic cancer, respectively. After de-N-glycosylation, relative quantitation of the hAGP glycans was carried out using stable isotope labeling and μZIC-HILIC-ESI-MS analysis. First, protein denaturing conditions prior to PNGase F digestion were optimized to achieve quantitative digestion yields, and the reproducibility of the established methodology was evaluated with standard hAGP. Then, the proposed method was applied to the analysis of the clinical samples (control vs. pathological). Pancreatic cancer samples clearly showed an increase in the abundance of fucosylated glycans as the stage of the disease increases and this was unlike to samples from chronic pancreatitis. The results gained here indicate the mentioned glycan in hAGP as a candidate structure worth to be corroborated by an extended study including more clinical cases; especially those with chronic pancreatitis and initial stages of pancreatic cancer. Importantly, the results demonstrate that the presented methodology combining an enrichment of a target protein by IAC with isotope coded relative quantitation of N-glycans can be successfully used for targeted glycomics studies. The methodology is assumed being suitable as well for other such studies aimed at finding novel cancer associated glycoprotein biomarkersPart of this study was supported by the Spanish Ministry of Science and Innovation (CTQ2011-27130 awarded to V. S-N and BIO 2010-16922 awarded to R.P.)Elsevier MassonMinisterio de Ciencia e Innovación (Espanya)infoinfo2015info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10256/12672http://hdl.handle.net/10256/12672© Analytica Chimica Acta, 2015, vol. 866, p. 59-68Articles publicats (D-B)reponame:Recercat. Dipósit de la Recerca de Catalunyainstname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)Inglésinfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.aca.2015.02.008info:eu-repo/semantics/altIdentifier/issn/0003-2670info:eu-repo/grantAgreement/MICINN//BIO2010-16922Tots els drets reservatsinfo:eu-repo/semantics/embargoedAccessoai:recercat.cat:10256/126722026-05-29T05:05:01Z
dc.title.none.fl_str_mv Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis
title Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis
spellingShingle Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis
Giménez, Estela
Pàncrees -- Càncer
Pancreas -- Cancer
Glicoproteïnes
Glycoproteins
Espectrometria de masses
Mass spectrometry
Marcadors tumorals
Tumor markers
title_short Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis
title_full Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis
title_fullStr Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis
title_full_unstemmed Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis
title_sort Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis
dc.creator.none.fl_str_mv Giménez, Estela
Balmaña, Meritxell
Figueras, Joan
Fort Martorell, Esther
Bolòs, Carme de
Sanz Nebot, María Victoria
Peracaula Miró, Rosa
Rizzi, Andreas
author Giménez, Estela
author_facet Giménez, Estela
Balmaña, Meritxell
Figueras, Joan
Fort Martorell, Esther
Bolòs, Carme de
Sanz Nebot, María Victoria
Peracaula Miró, Rosa
Rizzi, Andreas
author_role author
author2 Balmaña, Meritxell
Figueras, Joan
Fort Martorell, Esther
Bolòs, Carme de
Sanz Nebot, María Victoria
Peracaula Miró, Rosa
Rizzi, Andreas
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Ministerio de Ciencia e Innovación (Espanya)
dc.subject.none.fl_str_mv Pàncrees -- Càncer
Pancreas -- Cancer
Glicoproteïnes
Glycoproteins
Espectrometria de masses
Mass spectrometry
Marcadors tumorals
Tumor markers
topic Pàncrees -- Càncer
Pancreas -- Cancer
Glicoproteïnes
Glycoproteins
Espectrometria de masses
Mass spectrometry
Marcadors tumorals
Tumor markers
description In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [12C]- and [13C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. Human α1-acid-glycoprotein (hAGP) is an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation. However, it is not clear yet whether some particular glycans in hAGP can be used as biomarker for differentiating between these two pathologies. In this work, hAGP was isolated by immunoaffinity chromatography (IAC) from serum samples of healthy individuals and from those suffering chronic pancreatitis and different stages of pancreatic cancer, respectively. After de-N-glycosylation, relative quantitation of the hAGP glycans was carried out using stable isotope labeling and μZIC-HILIC-ESI-MS analysis. First, protein denaturing conditions prior to PNGase F digestion were optimized to achieve quantitative digestion yields, and the reproducibility of the established methodology was evaluated with standard hAGP. Then, the proposed method was applied to the analysis of the clinical samples (control vs. pathological). Pancreatic cancer samples clearly showed an increase in the abundance of fucosylated glycans as the stage of the disease increases and this was unlike to samples from chronic pancreatitis. The results gained here indicate the mentioned glycan in hAGP as a candidate structure worth to be corroborated by an extended study including more clinical cases; especially those with chronic pancreatitis and initial stages of pancreatic cancer. Importantly, the results demonstrate that the presented methodology combining an enrichment of a target protein by IAC with isotope coded relative quantitation of N-glycans can be successfully used for targeted glycomics studies. The methodology is assumed being suitable as well for other such studies aimed at finding novel cancer associated glycoprotein biomarkers
publishDate 2015
dc.date.none.fl_str_mv 2015
info
info
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10256/12672
http://hdl.handle.net/10256/12672
url http://hdl.handle.net/10256/12672
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.aca.2015.02.008
info:eu-repo/semantics/altIdentifier/issn/0003-2670
info:eu-repo/grantAgreement/MICINN//BIO2010-16922
dc.rights.none.fl_str_mv Tots els drets reservats
info:eu-repo/semantics/embargoedAccess
rights_invalid_str_mv Tots els drets reservats
eu_rights_str_mv embargoedAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier Masson
publisher.none.fl_str_mv Elsevier Masson
dc.source.none.fl_str_mv © Analytica Chimica Acta, 2015, vol. 866, p. 59-68
Articles publicats (D-B)
reponame:Recercat. Dipósit de la Recerca de Catalunya
instname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
instname_str Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
reponame_str Recercat. Dipósit de la Recerca de Catalunya
collection Recercat. Dipósit de la Recerca de Catalunya
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,81155

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