The Protein Core Facility (PCF) at CHOP addresses the overwhelming need for the technological resources to identify, produce and characterize new proteins. The PCF has four operating modules available: Protein expression, Characterization, Protein-Protein Interactions, and Proteomics. While many of our services are routine, we are also committed to collaborating with the CHOP community to overcome difficult research obstacles. We strongly encourage CHOP researchers to bring their research needs to our attention, so that we can try and provide solutions.
PROTEIN EXPRESSION AND PURIFICATION: With the advent of high-throughput DNA sequencing technology and the identification of an increasing number of genes and ORF's, there is an increasing demand for fast and efficient protein production. While some proteins can be expressed in bacteria, other proteins require other expression systems. These might include proteins that require certain chaperones for folding, are toxic to bacteria, contain multiple disulfides, have glycosylation sites, or require other extensive post-translational modifications for their activity. The PCF provides protein production in insect cells (sf9, sf21) using the Baculovirus expression system. We also offer assistance with expression of bacterial proteins or in vitro translation. For purification of the recombinant proteins we offer affinity, ion exchange and size exclusion chromatography.
PROTEIN CHARACTERIZATION: In addition to offering expression services, the PCF provides analyses such as mass spectrometry, gel electrophoresis, analytical and/or preparative chromatography, spectroscopy (fluorescence, circular dichroism, ultraviolet-visible), ultracentrifugation, and phosphorimaging. We also offer assistance in refolding recombinant proteins.
PROTEIN-PROTEIN INTERACTION: The identification of interacting protein partners is fundamental to the understanding of signaling networks and drug-discovery. The PCF has a number of tools for identifying new protein-protein interactions. Options include the SELDI protein-chip analyzer, analytical ultra-centrifugation, and Surface Plasmon Resonance (Biacore). We are also implementing, in collaboration with Invitrogen, an new protein-interaction array service. Once new partners are detected, we offer protein identification using mass spectrometry. Please consult with Steven Seeholzer for more information regarding these techniques.
PROTEOMICS: The PCF is committed to providinging the latest technologies in mass-spectrometry, multi-dimensional chromatography, protein-arrays etc. Utilizing these tools will enable us to uncover more new protein sequences and protein-protein partners so that we might have a better understanding of proteins within specific networks and at the center of pediatric diseases. This knowledge will be critical if we are to successfully understand and treat medical problems that arise at the biochemical level.