Unit: Protein Microsequencing and Analysis (Platform)
Director: d’Alayer Jacques
Protein Analysis and Microsequencing Laboratory continued its activities involving chemical protein microsequencing and analysis of proteins using ProteinChip technology. This new technology combines the capture of proteins on different chemical surfaces and their analysis by mass spectrometry.
These activities are for the benefit of laboratories both inside and outside the Pasteur Institute.
The laboratory is located at the " Génopole " building.
The protein microsequencing remained an important activity. Amino acid sequences are determined from purified or partially purified proteins and peptides.We can also obtain internal sequences by cutting proteins into fragments and purifiying them by HPLC chromatography before sequencing.
The amino acid sequences are performed by automatic sequencers using the Edman chemical degradation (two sequencers : ABI 473A and ABI 494 of Applied Biosystems are available). The laboratory also provides auxiliary support services, including advice about the microtechniques needed to prepare samples before sequencing.
350 samples were sequenced for 70 laboratories (including 36 from outside the Pasteur Institute).
The analysis of proteins using the ProteinChips Arrays technology of Ciphergen (www.ciphergen.com) is an expanding activity of our laboratory. The possibilities ot this technology (Protein capture on various chemical surfaces and analysis by mass spectrometry) are quite varied. For example : specific molecular interaction studies (discovery and validation of biological markers), phosphorylation studies, identification and characterization of proteins are only a part of the potential
applications. This analysis can be carried out with the very small amounts of biological material needed for mass spectometry.
In 2004, we performed analysis for 46 users.
Keywords: Protein, Microsequencing, ProteinChips, Ciphergen, Mass spectrometry