uu.seUppsala University Publications
Change search
ReferencesLink to record
Permanent link

Direct link
CIEF and MALDI-TOF-MS methods for analyzing forms of the glycoprotein VEGF165
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.
Show others and affiliations
2009 (English)In: Electrophoresis, ISSN 0173-0835, E-ISSN 1522-2683, Vol. 30, no 7, 1198-1205 p.Article in journal (Refereed) Published
Abstract [en]

The vascular endothelial growth factor (VEGF) is involved in different sicknesses (cardiovascular diseases, cancer, and other). Out of the many components of the VEGF family, the A splice variant with 165 amino acids (VEGF(165)) is the main component. in spite of the potential as biomarker that this protein has, information about its physico-chemical   characteristics is scarce. In this study CIEF and MALDI-TOF-MS methods for intact recombinant human VEGF(165) are developed and applied to analyze this glycoprotein expressed in glycosylating (Sf 21 insect cells) and non-glycosylating (Escherichia coli) systems. Different parameters influencing the CIEF separation were studied. The developed CIEF method allowed for the separation of up to seven peaks in the VEGF(165) expressed in insect cells and up to three in VEGF(165)  expressed in E. coli. The use of the presented method permits the estimation of the apparent pI of the different forms of VEGF(165)  expressed in insect cells to be in a range of 6.8-8.2. The three peaks with intermediate pI values are observed in the protein expressed in   both systems, insect cells and E. coli. The MALDI-TOF-MS method enabled to a rapid partial characterization of VEGF(165) based on its MS fingerprint. MALDI-MS analysis of VEGF(165) expressed in insect cells   shows the presence of, at least, four forms or groups of forms of VEGF(165) as a result of the different PTMs of the protein. According to the MALDI-MS analysis, VEGF(165) expressed in E. coli was produced   as a very homogeneous protein, although the results suggest the   existence of some PTMs in the protein. The patterns of VEGF(165) of   both origins obtained by CIEF and MALDI-MS indicate the possibility of using these analytical methods to compare samples from people with different pathophysiological conditions. This work is thus a starting point to make possible the study of the role of the various forms of VEGF(165) as biomarkers. Finally, to the best of our knowledge, this is the first time that intact VEGF(165) has been analyzed by CIEF and MALDI-TOF-MS.

Place, publisher, year, edition, pages
Weinheim: WILEY-VCH Verlag GmbH , 2009. Vol. 30, no 7, 1198-1205 p.
Keyword [en]
Angiogenesis, Biomarker, Glycoprotein, Isoform, VEGF
National Category
Chemical Sciences
Research subject
Analytical Chemistry
URN: urn:nbn:se:uu:diva-101421DOI: 10.1002/ELPS.200800592ISI: 000265478800014PubMedID: 19373805OAI: oai:DiVA.org:uu-101421DiVA: diva2:213037
Available from: 2009-04-27 Created: 2009-04-27 Last updated: 2011-03-30Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full textPubMed

Search in DiVA

By author/editor
Bergquist, Jonas
By organisation
Analytical Chemistry
In the same journal
Chemical Sciences

Search outside of DiVA

GoogleGoogle Scholar
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Altmetric score

Total: 397 hits
ReferencesLink to record
Permanent link

Direct link