The University of Queensland's (UQ) protein manufacture, preparation, analysis and stem cell research facilities are available through the Australian Institute for Bioengineering and Nanotechnology and School of Chemistry and Molecular Biosciences, both located on UQ's St Lucia campus.

Protein manufacture - Protein Expression Facility

Services

PEF delivers high quality recombinant proteins for discovery and preclinical research in diverse research areas for academic and industry organisations, globally and nationally. The protein production pipeline includes molecular cloning, expression optimisation, shake flask and bioreactor production, protein purification and characterisation. Each project commences with a tailored strategy for the target recombinant protein and its intended use.

Molecular cloning

Effective cloning strategy for construct generation

  • DNA template generation
  • Vector re-construction with tags for solubility, purification and detection
  • Site-directed mutagenesis

Expression

Tailored protein expression strategy for target protein and its intended use

  • Comprehensive expression systems in bacteria, yeast, baculovirus insect cell and mammalian cell systems 
  • Eukaryotic cell-free system for rapid protein analysis
  • Screening for optimal expression, solubility or secretion
  • Scale-up in shake flask and bioreactor or scale-down in plate format expression    

Purification

Tailored protein purification strategy for target protein and its intended use

  • Isolation of intracellular, membrane or secretory proteins
  • Analytical and preparative scale chromatography
  • Crossflow system for micro- and ultrafiltration
  • Fusion tag removal to restore native protein sequence
  • Endotoxin removal for animal studies

Characterisation

Biochemical and biophysical characterisation to ensure protein quality

  • Protein concentration and purity analysis 
  • Protein quaternary structure and aggregation analysis
  • Protein identification and sequencing 
  • Protein stability and formulation screening 
  • Protein complex and virus-like particle imaging
  • Protein complex and virus-like particle quantitative size distbriution
  • Endotoxin measurement

Equipment

PEF incorporates state-of-the-art facilities and a team of highly skilled specialists to enable a comprehensive and robust protein production pipeline.

General PEF equipment

  • 2 x Refrigerated benchtop centrifuge with swing bucket rotor, microplate rotor and fixed angle rotors 
  • NanoDrop spectrophotometer to quantify and qualify DNA, RNA and protein samples
  • Caliper LabChip GXII microfluidic instrument for high-throughput quantitation of DNA, RNA and protein samples
  • Multiskan GOTM microplate spectrophotometer for high-throughput optical density measurements and DNA, RNA and protein analysis
  • RVC 2-18 Rotional vacuum concentrator system

Molecular cloning

High throughput cloning for multiple construct generation

  • 4 x PCR Thermal cyclers
  • Nucleic acid electrophoresis systems

Expression

Shaking incubators for each expression system to support high-throughput parallel expression optimisation and preparative scale production for multiple proteins concurrently

  • 3-stacked shaking incubator (Bacteria)
  • 1 x shaking incubator (Yeast)
  • 3-stacked shaking incubator (Baculovirus/insect cell) 
  • 1 x shaking incubator (Mammalian cell)

Insect and mammalian cell analysis

  • Nikon eclipse TS100 microscope 
  • Countess automated cell counter

Preparative scale production and/or process development (up to 25 L) in bioreactor

  • 5 L UniVessel® BioStat B stirred-tank bioreactor (max. vol. 5 L Bacteria or Yeast)
  • 2 x 10 L UniVessel® BioStat® B stirred-tank bioreactor (max. vol. 20 L Bacteria or Yeast)
  • BioStat® RM 20/50 wave bioreactor (max. vol. 25 L Insect or Mammalian cells)

Purification

  • Avanti JXN-26 high-speed centrifuge
  • Branson sonicator with horn and microtip
  • APV-2000 Homogenizer
  • Sartocon® slice filtration sets 
  • 2 x AKTA Pure 25M chromatography system
  • 1 x AKTAxpress chromatography system

Characterisation

  • Protein gel electrophoresis chamber systems & iBlot 2 gel transfer device
  • Prometheus NT.48 for high-throughput label-free differential scanning fluorimetry (nanoDSF) technology to analyse protein aggregation, thermal stability and storage conditions
  • DynaPro plate reader for high-throughput dynamic light scattering (DLS) measurements of protein size distribution to determine protein aggregation, thermal stability and storage conditions
  • Agilent 1200 Series HPLC for analytical size exclusion chromatography to determine protein quaternary structure and aggregation
  • Asymmetrical-Flow Field-Flow Fractionation (AF4) to determine size distribution profile of virus-like particles (VLPs) and protein complexes
  • Endosafe®-PTSTM for measurement of endotoxin level

Resource centre

PEF provides quality assured materials to UQ researchers for molecular cloning, expression and purification.

  • Cell strains and cell lines
  • Plasmids
  • Enzymes (e.g. taq polymerase and proteases)

User equipment access

UQ researchers can be trained to access the following specialised equipment: 

  • UniVessel® BioStat B stirred-tank bioreactor
  • BioStat® RM 20/50 wave bioreactor
  • Prometheus NT.48
  • Caliper labchip GXII

Training

PEF provides consultation and training to upskill UQ staff and students (Honours, Masters and PhD) in all aspects of recombinant protein production.

  • Consultation
  • Workshops  

Symposium

PEF has hosted Protein Production Symposiums to provide network opportunities with local/interstate/international researchers in the field and share key technologies and strategies for protein production. 

PEF liaison with other research facilities

  • AGRF
  • Mass Spectrophotmetry Facility 
  • Centre for Microscopy & Microanalysis (CMM)

Contact

Australian Institute for Bioengineering and Nanotechnology

(07) 3346 3979

Australian Institute for Bioengineering and Nanotechnology, AIBN Building (Building 75), St Lucia campus

Protein manufacture - National Biologics Facility

Services

The National Biologics Facility was established through the NCRIS programme to assist academic and industrial researchers to bridge the gap between research discoveries and the bioprocesses required to produce material for pre-clinical studies.  The Facility produces high-quality recombinant proteins, with a focus on biologics for pre-clinical studies. The NBF specialises in use of mammalian cell expression technology for proteins with potential therapeutic or commercial uses.  

Equipment

Activities and services include:

  • Protein and Antibody Engineering
  • Antibody discovery through phage display
  • Molecular Biology
  • Transient expression of proteins in CHO and HEK cells
  • Stable cell line development, creating cell lines suitable for GMP manufacture of product
  • Upstream bioprocessing
  • Downstream bioprocessing
  • Protein purification
  • Protein analytics
  • Access to instrumentation such as Isothermal Colorimetry and Surface Plasmon Resonance

Contact

Doctor Martina Jones

Australian Institute for Bioengineering and Nanotechnology

(07) 3346 3178

Australian Institute for Bioengineering and Nanotechnology, AIBN Building (Building 75), St Lucia campus

Protein preparation and analysis

Services

Facility houses a range of equipment to facilitate sample preparation and analysis of proteins and other biological molecules, including scanning of gels, fractionation / purification of proteins, analysis of protein-protein and other molecular interactions.

Equipment

  • Typhoon FLA7000 phosphor Imager
  • Beckman DTX880 Multimode plate reader
  • Nano-ITC Microcalorimeter
  • GE Ai600 Imager (chemiluminescence, RGB LED fluorescence, UV, Light)
  • Typhoon Variable Mode Scanner (Phospho-storage, chemiluminescence and fluorescence)
  • IPGphor iso-electric focussing units (7, 13 and 24 cm IEF strips)
  • Ettan Dalt II 2nd dimension Electrophoresis System and Gel Caster (24 cm)
  • Progenesis SameSpots software for single and multi-stained gels
  • Waterbath Sonicator
  • Thermo Savant: Speedvac and Freeze Dryer
  • Odyssey Classic Infra-Red laser Imager  
  • Odyssey CLx infra-Red laser Imager
  • SpectraMAX 190 Plate Reader (U/V vis)
  • SpectraMAX 190 Plate Reader 
  • SpectraMAX 250 Plate Reader
  • CLARIOstar Multimode plate reader with environmental control (luminescence, monochromatic fluorescence and absorbance)
  • Cytofluor fluorescence plate reader
  • Eppendorf refrigerated bench top microcentrifuge
  • Umax Image scanner for transmittance scanning of film and gels.
  • Agilent Bioanalyser: for RNA, DNA and protein spectral imaging
  • Microcal iTC200 microcalorimeter
  • Biacore 3000
  • Beckman Multimode detector
  • Beckman Optima XL/I analytical ultracentrifuge
  • AKTA fplc and fraction collector
  • AKTA Pure and fraction collector
  • Beckman Analytical Ultracentrifuge
  • Agilent 1100/1200 capillary HPLC system with UV detector and fraction collector
  • Shimadzu Prominence HPLC system with UV and fluorescence detectors, autosampler and fraction collector
  • Jasco Circular Dichroism (CD) spectrometer 

Contact

Doctor Amanda Nouwens

School of Chemistry and Molecular Biosciences

(07) 3346 9490

Doctor Steven Mason

School of Chemistry and Molecular Biosciences

(07) 3365 4872

Molecular Biosciences Building 76-409, St Lucia campus

Stem cell research

Services

The Pluripotent Stem Cell Core Facility facilitates pluripotent stem cell research both within the University of Queensland and throughout Australia. these services include provision of Human Embryonic Stem Cell Lines (hESCs), Mouse Embryonic Fibroblasts (MEFs), Custom Generated Induced Pluripotent Stem Cells (iPSCs), Genetically Engineered iPSCs Using CRISPRs, Comprehensive Training in Generation and Maintenance of iPSCs, and Mycoplasma, Teratoma & Karyotype Assays.

Equipment

N/A

Contact

Doctor Eva Thoma

Australian Institute for Bioengineering and Nanotechnology

(07) 3365 0339

Australian Institute for Bioengineering and Nanotechnology, AIBN Building 75, St Lucia campus