Université de Strasbourg
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Chimie bioanalytique

Teachers :   Pr Laurence Sabatier

                   Laboratory of Bioorganic Mass Spectrometry (LSMBO)

                   Analytical Sciences Department, IPHC

                   E-mail: laurence.sabatier@unistra.fr

                   Dr Véronique Delval

                   Laboratory of Bioorganic Mass Spectrometry (LSMBO)

                   Analytical Sciences Department, IPHC

                   E-mail: veronique.delval@unistra.fr

Duration: 21h

 

EXPECTED SKILLS:

- Acquire an expertise in the purification methods adapted to biomolecules and in the use of mass spectrometry for their characterization.

- Understand the advantages and drawbacks of the different analytical approaches for the biomolecules.

- Advise and design a convenient proteomic strategy.

- Handle and interpret data.

- Integrate the results from different analyses

- Manage a bioanalytical project

CLASS CONTENTS :

  • · Recall on the structure and the properties of biomolecules (focused on those useful for their purification)
  • · Purification of biomolecules using liquid chromatography: size exclusion, ion exchange, chromatofocalisation, hydrophobic interaction HIC, reverse phase, hydrophilic interaction HILIC, metal chelation, affinity. Stationary phases/mobile phases/ setting up of purification conditions. Preparative scale and miniaturized scale: constraints, instrumental modifications and applications.
  • · Purification of biomolecules using monodimensional and bidimensional electrophoresis. Denaturating and native conditions. Principles, parameters to optimize (sample preparation, extraction, solubilisation, depletion, minor proteins enrichment) and applications. Off gel electrophoresis.
  • · Contribution of mass spectrometry to the characterization of biomolecules: ionization sources for biomolecules, interpretation of mass spectra MS and MS/MS of biomolecules, identification of a protein from an electrophoresis gel or a chromatographic fraction, mass fingerprint, de novo sequencing, characterization of post-translational modifications.
  • · Shotgun and targeted proteomic strategies
  • · Peptidomic and proteomic differential analyses
  • · Strategies of relative and absolute quantification of proteins in complex mixtures.
  • · Supramolecular mass spectrometry
  • · Characterization of protein interactions

BIBLIOGRAPHY :

- Principles and Practice of Bioanalysis, 2nd Edition (2008), Richard F. Venn,  Publisher : CRC Press

- Bioanalytical chemistry (2004), A. Manz, N. Pamme, D Lossifidis, Editeur : Imperial College Press

- Mass spectrometry of proteins and peptides : methods and protocols , 2nd Edition (2008), M. S. Lipton & L. Pasa-Tolic, Publisher: Humana Press

- Principles of Mass Spectrometry Applied to Biomolecules, (2006), Chava Lifshitz and Julia Laskin, Publisher: Wiley

- Principles of Proteomics, 2nd edition (2013), Richard Twyman, Eds Garland Science