CEA Direction des sciences du vivant : ProteoLab and BioChipLab:Integrated microsystems for preparation of protein sample for mass spectrometry identification

ProteoLab and BioChipLab:Integrated microsystems for preparation of protein sample for mass spectrometry identification

Biochips at the CEA
Biochips at the CEA
A MicroArray Production site in Evry
The european MeDICS project: a miniaturized cell sorter
Multi-Patch: a chip-based ion-channel assay system for automatic screening
Phenotypic analysis on a chip
Towards high performance genomic and proteomic analyses
ProteoLab and BioChipLab:Integrated microsystems for preparation of protein sample for mass spectrometry identification

Aim
ProteoLab and BioChipLab are innovative microsystem interfaces designed to increase sensitivity of protein identification by mass spectrometry. They offer 3 major advantages
- higher yields and faster kinetics of protein digestion
- low sample volumes
- direct interfacing with the mass spectrometer with no dead volumes or sample loss.

Principle
Sample preparation is carried out in 2 steps:
1 Proteins (isolated from a 2D gel separation) are digested by trypsin (ie cut into smaller peptides), grafted to the surface of the microreactor.
2 Peptides are separated by on chip chromatography and sent to the mass spectrometer for identification.

A modular architecture
The proteolysis and separation modules are designed for optimum fluid flow profiles and maximum surface to volume ratios. Fluid transport is controlled by electro-osmotic flow. Interface with the mass spectrometer is achieved by electrospray injection.

Separation pillars
Specific proteolysis microreactor



		MS ElectroSpray Interface
Electrokinetic liquid pumps

First prototypes
First prototypes were microfabricated by the CEA/Leti on silicon. The Trypsin protease was immobilized on the pillars of the first module. Sample injection and recovery is done by capillary inlet/outlet ports through the elastomer cover.

Wafer 100 mm	Detail of pre-concentration module	Separation module with mixer
Wafer 100 mm	Detail of separation column

Partners
CEA:
Microtechnology and Electrokinetics
Biological and Chemical Integration
Research Models and Biological Validation
USTL and CNRS:
Microtechnology and Electrospray Output Interface In situ Polymer Chemistry
Osmooze: Droplet Electrotransport
Sanofi-Synthélabo: Industrial Models and Validation.

Financial support
Protéolab: Conseil Régional Rhône Alpes.
BioChipLab: MEFI - Ministère de l'Economie, des Finances et de l'Industrie.

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Contacts:
Leti: Pierre Puget
Life Sciences Division: Jérôme Garin (jerome.garin@cea.fr)