wall PROTEOMICS spear METABOLOMICS snake VENOMICS tree GENOMICS rope GLYCOMICS fan PHENOMICS Fluorescence Bringing Life to Proteins and Proteins to Life FABLS Network Launch Feb 2005

Australian Proteome Analysis Facility Ltd (APAF) birthplace of the word “proteomics” in 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 (1DE, 2DE, IPG, native gel, blot) for : 1.Protein 2.Enzymatic activity 3.Post-translational modification (PTM) 4.Peptide bioactives

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 APAF’S MAJOR ACTIVITIES

Using Fluorescence to Turn Proteomic Bottlenecks into Development Opportunities Removal of 99.9% most abundant human biofluid proteins & from other biological sources Improved sensitivity via high throughput sample processing direct on biochip technology Development of new improved fluorescent all protein stains Development of fluorescent spot cutter/digestion robot MS/MS capability direct off-chip MS/MS capable affinity chips Development of less operator intensive HTS imaging systems Proteomic LIMS development Development of 2D-LC HTS Fractionation of relevant proteins

hemopexin HSA fibrinogen IgG heavy chain  Ig light chain ApoA1 transferrin a1-antitrypsin haptoglobin leucine-rich a2 glycoprotein A2 HS-glycoprotein fetuin transthyretin a2 M fibrinogen 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

SarA D C B A IsaA D B C A Identifying S. aureus proteins associated with methicillin resistance Meth sensitive TXMeth resistant COL COL +TX

New fluorescent stains for deeper drilling of the proteome 250ng 30ng 4ng More sensitive detection means more proteins of interest

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, AnchorChips, SELDI & STS biochips Apply Sample Dry Sample STS Biochip Bruker AnchorChip Dry Sample Apply Sample Apply Sample ABI 4700 Opti-TOF MALDI Plate Apply Sample Wash Sample & Dry SELDI Wash Sample Focus & Dry Diffuse Sample Over Large Area Diffuse Sample Over Large Area Selective Binding Non-selective Binding Concentrate Sample Concentrate Selected Sample

Chemical proteomics: active site directed probes Leung, D.; Hardouin, C.; Boger, D. L.; Cravatt, B. F. Nat. Biotech. 2003, Serine proteases Discovery of known and unknown serine hydrolase activity

TGR’s SureFire TM Technology Removing a Bottleneck in drug / bioactive discovery High throughput, Sensitive, Quantitative Assays

Thanks from all the APAF team