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„Aurel Vlaicu” din Arad


Project director: Dr. Corina Flangea


Subdomain: LS2.Genetics, Genomics, Bioinformatics and Systems Biology: genetics, population genetics, molecular genetics, genomics, transcriptomics, proteomics, metabolomics, bioinformatics, computational biology, biostatistics, biologicalmodelling and simulation, system

Main Research Area: LS2_5.Glycomics

Secondary Research Area: LS2_3.Proteomics

Project title in English: Glycomics of brain glycosaminoglycans by advanced mass spectrometric and related hyphenated techniques

Project title in Romanian: Glicomica glicozaminoglicanilor din creier prin tehnici avansate de spectrometrie de masa si tehnici complementare

Project duration: 36 month (01.10.2011-30.09.2014)

Budget: 750.000 lei

Financial support: Romanian National Authority for Scientific Research, CNCS-UEFISCDI (Consiliul National al Cercetarii Stiintifice - Unitatea Executiva pentru Finantarea Invatamantului Superior, a Cercetarii, Dezvoltarii si Inovarii)

Contract No: 93/05.10.2011

Research team:

  • Corina Flangea - Project director
  • Alina Zamfir - Senior researcher 
  • Catalin Schiopu - PhD student

Project Summary

Chondroitin sulfate (CS) and dermatan sulfate (DS) glycosaminoglycans (GAG) are covalently linked to core proteins, forming proteoglycans (PG) with a wide distribution in extracellular matrix of brain. Recent studies have shown that proportion of CS/DS disaccharide units differing in the degree and profile of sulfation are involved in the functional diversity of neurons; however, so far not detailed structural analysis of brain GAGs was pursued. To close this gap, we want to introduce for the first time in glycomics of brain GAGs, an advanced mass spectrometry (MS)-based system based on fully automated chip nanoelectrospray ionization (nanoESI) and multistage MS (MSn) for structural investigation up to decasaccharides CS/DS chains regarding the sulfation pattern. The strategy requires the combination of biochemical tools and an analytical platform. CS/DS chains released by beta-elimination are purified and depolymerized with chondroitin AC lyase in parallel with chondroitin B lyase. The mixture of chains with different size length will be separated on size exclusion chromatography (SEC). The collected fractions from SEC are automatically infused into MS by chip-nanoESI. The species with the same length but different sulfation content are screened and isolated for MSn experiments. MSn technique is able to provide fine structural information about sulfate content and to determine for the first time the position of sulfate group inside the monomer ring for longer CS/DS chain.


Articles in impact factor journals

1. Flangea C, Petrescu AJ, Seidler DG, Munteanu CV, Zamfir AD. Identification of an unusually sulfated tetrasaccharide chondroitin/dermatan motif in mouse brain by combining chip-nanoelectrospray multistage MS(2) -MS(4) and high resolution MS.

Electrophoresis, 2013 34, 1581-1592; impact factor: 3.30

2. Flangea C, Fabris D, Vukelić Ž, Zamfir AD. Mass spectrometry of gangliosides from human sensory and motor cortex

Aus. J. Chem., 2013, 66, 781-790; impact factor: 2.34

3. Zamfir AD, Fabris D, Capitan F, Munteanu C, Vukelić Ž, Flangea C. Profiling and sequence analysis of gangliosides in human astrocytoma by high resolution mass spectrometry

Anal. Bioanal. Chem., 2013, 405, 7321-7355 impact factor: 3.77

4. Flangea C, Sisu E, Seidler DG, Zamfir AD. Analysis of oversulfation in biglycan chondroitin/dermatan sulfate oligosaccharides by chip-based nanoelectrospray ionization multistage mass spectrometry

Anal Biochem., 2012, 420, 155-162; impact factor: 2.96

5. Sarbu M, Robu A, Peter-Katalinic J, Zamfir AD. Automated Chip- nanoelectrospray mass spectrometry for Glycourinomics in Schindler Disease type I.

Carbohydr. Res. 2014, 398, 90-100; impact factor: 1.96

6. Sarbu M, Ghiulai RM, Zamfir AD. Recent developments and applications of electron transfer dissociation mass spectrometry in proteomics.

Amino Acids. 2014, 46, 1625-1634; impact factor: 3.65 


Book chapters

1. Zamfir AD, Flangea C, Serb A, Zagrean AM, Rizzi AM, Sisu E. Separation and identification of glycoforms by capillary electrophoresis with electrospray ionization mass spectrometric detection.

Methods Mol Biol., 2013, 951, 145-169

2. Zamfir AD, Flangea C, Serb A, Sisu E, Zagrean L, Rizzi A, Seidler DG. Brain chondroitin/dermatan sulfate, from cerebral tissue to fine structure: extraction, preparation, and fully automated chip-electrospray mass spectrometric analysis.

Methods Mol Biol., 2012, 836, 145-159

3. Zamfir AD. Nanochips for Mass Spectrometry and Applications in Biomedical Research in: "Research in horizons of nanosystems structure, properties and interactions" Apple Academics, Ontario-New Jersey, Canada-USA, pp.386-443; 2014

4. Sirbu M, Zamfir AD. Nanofluidics-Based Mass Spectrometry. Applications for Biomarker Discovery in Lysosomal Storage Diseases in:
"Nanoparticles’ Promises and Risks,"

Springer International Publishing Switzerland, pp. 137-165; 2014

Conference presentations

1. C. Schiopu, F. Capitan, Ž.Vukelić, C. Flangea, A. D. Zamfir

Chip-nanoelectrospray quadrupole time-of-flight tandem mass spectrometry of human meningioma glycolipids

46th German Society for Mass Spectrometry Conference, March 10-14, 2013, Berlin, Germany

2. A.D. Zamfir

Clinical applications of chip-based mass spectrometry

2nd Middle Eastern and Mediterranean Sea Region Countries Mass Spectrometry Conference, June 30 - July 4, 2013, Siena, Italy

3. C. Schiopu, F. Capitan, A. Serb, Ž. Vukelić, E. Sisu, A.D. Zamfir.

Molecular expression and structure of gangliosides in brain metastasis of lung adenocarcinoma by NanoMate-QTOF MS and MS/MS

Joint Conference of Polish Mass Spectrometry Society and German Mass Spectrometry Society, March 4-7 2012, Poznan, Poland

4. A.F. Serb, S. Dragan, C. Flangea, E. Sisu, A.M. Zagrean, Ž. Vukelić, A.D. Zamfir. Application of Modern Mass Spectrometric Methods in Neuroscience Research: Mapping and Sequencing of Human Brain Gangliosides.

Joint Conference of Polish Mass Spectrometry Society and German Mass Spectrometry Society, March 4-7 2012, Poznan, Poland

5. A.D. Zamfir, C. Flangea, R.M. Ghiulai, D.G. Seidler.

Identification and structural analysis by chip-based nanoelectrospray mass spectrometry of novel brain-associated chondroitin/dermatan sulfate motifs

International Mass Spectrometry Conference, September 11-22 2012, Kyoto, Japan

6. C. Flangea, E. Sisu, A.D. Zamfir.

Chip-based nanoelectrospray ionization mass spectrometry for biomarker discovery in brain

The 3rd Conference of the National Neuroscience Society of Romania with CEERC-IBRO international Session, October 18-19 2012, Bucharest, Romania

The paper was awarded the 3rd Prize of the Jury

7. C. Flangea, C. Schiopu, F. Capitan, D. Seidler, E. Sisu, A.D. Zamfir.

Determination of decorin chondroitin/dermatan sulfate expression in human embryonic kidney cells by fully automated chip-based nanoelectrospray mass spectrometry.

International Symposium Research and Education in Innovation Era, November 08-09 2012, Arad, Romania

8. C. Flangea, A.D. Zamfir.

Identification of novel chondroitin/dermatan sulfate domain in brain by chip-nanoelectrospray multistage and high resolution mass spectrometry.

5th International Congress and the 31th Annual Session of the Romanian Society for Cell Biology, June 05-09 2013, Timisoara, Romania

9. Cristina Mosoarca, Loredana Lupu, Iosif Malaescu, Alina D. Zamfir
Top-down analysis by electron transfer/collision induced dissociation of noncovalent protein-ganglioside complexes

25th Mass Spectrometry Forum, February 17-19 2014, Viena, Austria.


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