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Development of a CE-MS method to analyze components of the potential biomarker vascular endothelial growth factor 165
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Physical and Analytical Chemistry, Analytical Chemistry.
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Physical and Analytical Chemistry, Analytical Chemistry.
2009 (English)In: Electrophoresis, ISSN 0173-0835, E-ISSN 1522-2683, Vol. 30, no 13, 2355-2365 p.Article in journal (Refereed) Published
Abstract [en]

The vascular endothelial growth factor 165 (VEGF(165)) is the predominant form of the complex VEGF-A family. Its angiogenic effect is   involved in many physiological and pathological events. For this   reason, its roles as a potential biomarker and as a therapeutic drug   have been considered. Nevertheless, very little is known about the   existence of different forms of VEGF(165) arising from glycosylation  and potentially from other PTMs. This aspect is important because  different forms may differ in biological activity (therapeutic drug   application) and the pattern of the different forms can vary with   pathological changes (biomarker application). In this work a CE-MS   method to separate up to seven peaks containing, at least, 19 isoforms   of intact VEGF(165) is described. Comparison between human VEGF(165)   expressed in a glycosylating system, i.e. insect cells, and in a   non-glycosylating system, i.e. E. coli cells, has been carried out. The   method developed provides structural information (mass fingerprint)   about the different forms of VEGF(165) and after the deconvolution and   the analysis of the MS spectra, PTMs pattern of VEGF(165) including   glycosylation. and loss of amino acids at the N- and C-terminus was   identified. Glycans involved in PTMs promoting different glycoforms   observed in the CE-MS fingerprint were confirmed by MALDI-MS after   deglycosylation with peptide N-glycosidase F. This approach is a   starting point to study the role of VEGF(165) as a potential biomarker   and to perform quality control of the drug during manufacturing. To our   knowledge this is the first time that a CE-MS method for the analysis  of VEGF(165) has been developed.

Place, publisher, year, edition, pages
Weinheim: Wiley-VCH Verlag GmbH , 2009. Vol. 30, no 13, 2355-2365 p.
Keyword [en]
CE, Polyamine coating, PTMs, Time of flight MS, Vascular endothelial growth factor
National Category
Chemical Sciences
Research subject
Analytical Chemistry
Identifiers
URN: urn:nbn:se:uu:diva-113915DOI: 10.1002/elps.200800738ISI: 000268626900015PubMedID: 19621363OAI: oai:DiVA.org:uu-113915DiVA: diva2:292132
Available from: 2010-02-04 Created: 2010-02-04 Last updated: 2017-12-12Bibliographically approved

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