Fluorescence Bringing Life to Proteins and Proteins to Life wall PROTEOMICS fan PHENOMICS spear METABOLOMICS snake rope GLYCOMICS tree GENOMICS FABLS Network Launch Feb 2005 VENOMICS
Australian Proteome Analysis Facility Ltd (APAF) • birthplace of the word “proteomics” in 1995 • 1 st medium: high-throughput proteomics Facility in 1996 • wholly-owned public company in 2002 • Major National Research Facility until 2006 • world class “one-stop” proteomics discovery facility with 40 EFTS located in 4 purpose-built centres of excellence that work synergistically • service provider, discovery partner, educator, collaborator & proteomics technology developer
Australian Proteome Analysis Facility Ltd (APAF) Discovery, development & utilisation of sensitive, accurate & cheap fluorescence proteomics probes (1 DE, 2 DE, IPG, native gel, blot) for : 1. Protein 2. Enzymatic activity 3. Post-translational modification (PTM) 4. Peptide bioactives
APAF’S MAJOR ACTIVITIES • Client-based Contract Research & Life Sciences Collaborations • Linkages with other MNRFs • Technology E, R&D • Education & Training Program • In-house Scientific Programs • • Biomarker Discovery Invasive Cancer Biology Agricultural Productivity Traits Bacterial Pathogenesis
Using Fluorescence to Turn Proteomic Bottlenecks into Development Opportunities Removal of 99. 9% most abundant human biofluid proteins & from other biological sources Fractionation of relevant proteins Development of new improved fluorescent all protein stains Development of less operator intensive HTS imaging systems Development of 2 D-LC HTS Development of fluorescent spot cutter/digestion robot Improved sensitivity via high throughput sample processing direct on biochip technology MS/MS capability direct off-chip MS/MS capable affinity chips Proteomic LIMS development
a 2 M Ig. G heavy chain g hemopexin A 2 HS-glycoprotein fetuin HSA transferrin fibrinogen a 1 -antitrypsin Ig. G heavy chain g leucine-rich a 2 glycoprotein haptoglobin Apo. A 1 Ig light chain transthyretin Drilling deeper into the plasma proteome by removal of high abundance proteins
PLASMA PROTEINS NEEDING TO BE VISUALISED FOR BIOMARKER DISCOVERY VISUALISATION REQUIRED TO DISCOVER NOVEL PLASMA PROTEIN DISEASE BIOMARKERS
Identifying S. aureus proteins associated with methicillin resistance A B Sar. A Meth sensitive 8325 A C Sar. A 8325 +TX Isa. A Meth resistant COL B Isa. A Sar. A COL +TX Isa. A C D D Isa. A
250 ng New fluorescent stains for deeper drilling of the proteome More sensitive detection means more proteins of interest 30 ng 4 ng
WHAT’S HOT IN PROTOEMICS !!! APAF and LCI STS Biochip R&D program AIM To develop the new STS biochip platform for the rapid affinity capture, concentration and TOF/TOF identification of peptides and proteins
MALDI, Anchor. Chips, SELDI & STS biochips Dry Sample Apply Sample ABI 4700 Opti-TOF MALDI Plate Bruker Anchor. Chip Non-selective Binding Dry Sample Apply Sample Non-selective Binding Apply Sample STS Biochip Concentrate Sample Wash Sample & Dry Selective Binding SELDI Diffuse Sample Over Large Area Wash Sample Focus & Dry Selective Binding Diffuse Sample Over Large Area Concentrate Selected Sample
Chemical proteomics: active site directed probes Serine proteases Discovery of known and unknown serine hydrolase activity Leung, D. ; Hardouin, C. ; Boger, D. L. ; Cravatt, B. F. Nat. Biotech. 2003, 687 -71.
TGR’s Sure. Fire Technology TM Removing a Bottleneck in drug / bioactive discovery High throughput, Sensitive, Quantitative Assays
Thanks from all the APAF team
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