Unit: Protein Microsequencing and Analysis

Director: d'Alayer Jacques

During 2003 the Protein Microsequencing and Analysis Platform continued its activities involving chemical protein microsequencing and analysis of proteins using ProteinChipTM 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.

In July, the laboratory was relocated to the Genopole building.

The chemical protein microsequence remained the Platform's principal 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 sequence are determined by automatic sequencers using the Edman chemical degradation technique (two sequencers : ABI 473A and ABI 494 of Applied Biosystems are available). The Platform also provides auxiliary support services, including advice about the micro-techniques needed to prepare samples before sequencing and aid in interprétation of data obtained.

During 2003, 53 laboratories, including 31 from outside the Pasteur Institute, have used our protein microsequencing service (> 410 samples were run).

The analysis of proteins by mass spectrometry (SELDI-TOF) after capture on various chemical surfaces (ProteinChip Arrays) using the Ciphergen's technology (www.ciphergen.com) is an expanding activity of our laboratory. A large number of diverse analyses can be carried out : specific molecular interaction studies (discovery and validation of biological markers), purification, identification and characterization of proteins, and phosphorylation studies are only a part of the potential applications.

These analyses can be carried out with the very small amounts of material needed for mass spectrometry.

In 2003, 30 laboratories have used our Ciphergen based protein analysis service.

The laboratory participated in training and seminars.

Keywords: Protein, Microsequencing, ProteinChip, Ciphergen, Mass spectrometry

Activity Reports 2003 - Institut Pasteur

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