Unit: Protein Microsequencing and Analysis (Platform)
Director: d’Alayer Jacques
Protein Analysis and Microsequencing Platform continued its activities involving chemical protein microsequencing and protein analysis using ProteinChip technology. This technology combines the capture of proteins on different chemical surfaces and their analysis by mass spectrometry (SELDI-TOF-MS).
These activities are for the benefit of laboratories both inside and outside the Pasteur Institute.
The chemical protein sequencing remainded its principal activity. Amino acid sequences are determined from purified or partially purified proteins or peptides. We can also obtained internal sequences by cleaving proteins into fragments and purifying them by HPLC chromatography before sequencing.
Amino acid sequences are performed by automatic sequencers using the Edman chemical degradation (two sequencers are available : ABI 473A and ABI 494 from Applied Biosystems). The laboratory also provides auxiliary support services, including advice about the microtechniques needed to prepare samples before sequencing.
380 samples were sequenced for 67 laboratories (including 32 from foreign countries).
The analysis of proteins using the ProteinChips Arrays technology of Ciphergen (www.ciphergen.com) is an expanding activity in our laboratory. The possibilities of 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 very small amounts of biological material needed for mass spectrometry (a few picomoles).
In 2005, we performed analysis for 36 users. Hands-on trainings were organized
Keywords: Protein, Microsequencing, ProteinChips, Ciphergen, Mass spectrometry