Capabilities

Capabilities

One of APAF’s goals is to continually push the limits of these technologies to answer the most challenging research questions. This ensures that Australian researchers in the life sciences, medical, agriculture, food and biotech sectors will continue to have access to the latest generation of advanced technologies, and thereby retain global leadership across their respective fields.

APAF’s multidisciplinary team and wide range of instrumentation allow us to serve the community across diverse research disciplines and provide a range of services:

Amino Acid Analysis (AAA)

  • Instruments available:
    • Premier Class, I-class, Acquity Class UPLC instruments
  • General Information:
    • Amino acid analysis is a technique to determine the quantities of individual amino acids, the amino acid ratio or composition, and is regarded as the gold standard for protein and peptide quantification. Colorimetric dye-binding assays are often used to determine protein concentration but these assays can be inaccurate and vary from protein to protein.
    • AAA is extensively applied to provide nutrition data, in quality control, for protein quantitation and in research applications.
    • AAA can be applied to a wide variety of sample types, from foodstuffs to plasma, and from synthetic peptides to dietary supplements.
    • Outcomes of AAA include the amounts of individual amino acids present in a sample (in mg/g, or µg/g) and their molar ratios.
    • For detailed information click here
  • Accessible for fee-for-service work
  • Contact: Dr. David Cantor – david.cantor@mq.edu.au
  • NATA Accredited: Yes - refer to the Scope of Accreditation 20344 for further details

Multiplexed Immunoassay (MIA)

  • Instruments available:
    • Bioplex 200, Plate-washing station and Andrew+ Pipetting Robot
  • General Information:
    • Multiplex immunoassays measure up to 65 different analytes in a single reaction. This is particularly beneficial for precious samples, such as plasma or niche biofluids, or when only a small volume is collected for analysis.
    • MIA is predominately performed for measuring concentrations of cytokines, chemokines, growth factors, and phosphoproteins. There are many human assays commercially available for a variety of physiological states such as cancer biomarker panels, diabetes, metabolic, inflammation, cell signalling, etc, as well as for a non-human species (i.e., mouse, rat, canine).
    • Outcomes of MIA include the simultaneous, absolute quantitation of analytes present in a sample (in pg/mL, or ng/mL).
  • Accessible for training and student use, or fee-for-service work
  • Contact name: Dr. David Cantor – david.cantor@mq.edu.au
  • NATA Accredited - refer to the Scope of Accreditation 20344 for further details

Enzyme-linked Immunosorbent Assay (ELISA)

  • Instruments available:
    • Fluostar Omega and Andrew+ Pipetting Robot
  • General Information:
    • ELISA assay is considered as the gold standard colorimetric immunoassay assay for detecting and quantifying specific analytes in complex biological samples. ELISA has been used as a diagnostic tool in medicine, clinical trials, food allergens, cell culture, plant pathology, and quality-control check in various applications
    • Outcomes of ELISA are the absolute quantitation of a given analyte within a sample or extract (in pg/mL, or ng/mL)
  • Accessible for training and student use, or fee-for-service work
  • Contact name: Dr. David Cantor – david.cantor@mq.edu.au

High Performance Liquid Chromatography (HPLC)

  • Instruments available – Agilent 1200/1260/1290 Infinity series
  • General Information:
    • APAF offers a variety of HPLC methods for separation and analysis of proteins and their fragments, based on their size, hydrophobicity and/or charge. APAF offers additional HPLC services such as high-pH fractionation and size-exclusion chromatography.
    • HPLC is being used for separation or semi purification of proteins, monitoring batch to batch protein purification and identifying post-translational modifications of proteins in the field of molecular biology, genetics, microbiology, biochemistry and clinical chemistry.
    • Size-exclusion chromatography is generally used to separate biological molecules, to determine molecular weight distributions of proteins and peptides.
    • Outcomes of HPLC depend largely upon the experimental design. These can include the identification and relative quantification of particular proteoforms, determination of molecular weight distribution or sample fractionation. Methods of interest include:
    • A1/A2 β-casein quantitation in milk
    • Lactoferrin analysis from nutraceutical products
  • Accessible for training and student use, or fee-for-service work
  • Contact name: Dr. David Cantor – david.cantor@mq.edu.au
  • NATA Accredited - refer to the Scope of Accreditation 20344 for further details

Gel electrophoresis

  • General Information:
    • Gel electrophoresis is used for separation or semi purification of proteins, monitoring batch to batch protein purification, identifying post-translational modifications of proteins, and validation of putative biomarkers by Western blot analysis in the field of molecular biology, genetics, microbiology, biochemistry and clinical chemistry.
  • Accessible for training and student use, or fee-for-service work
  • Contact name: Dr. David Cantor – david.cantor@mq.edu.au
  • NATA Accredited - refer to the Scope of Accreditation 20344 for further details

Protein identification and sequence coverage

  • General Information:
    • Proteins are digested into peptides and analysed by LC-MS. Samples can be from gel pieces, isolated fractions from LC experiments, or other preparations. Generated spectra are searched against protein databases, and all peptides and proteins identified are reported, and sequence coverage of proteins of interest can be calculated and reported.
  • Accessible for training and student use, or fee-for-service work
  • Contact name: Dr. Muhammad Zenaidee - muhammad.zenaidee@mq.edu.au
  • NATA Accredited - refer to the Scope of Accreditation 20344 for further details

Relative protein quantification

  • General Information:
    • Protein from cell lysates, tissue sample or other biological materials are digested into peptides and analysed by LC-MS. Generated spectra are searched against protein databases for protein identification, and relative protein abundance across samples calculated. Protein abundance is given for all quantified proteins, and differentially expressed proteins identified. Labelled and unlabelled experiments can be facilitated.
  • Accessible for training and student use, or fee-for-service work
  • Contact name: Dr. Muhammad Zenaidee - muhammad.zenaidee@mq.edu.au
  • NATA Accredited - refer to the Scope of Accreditation 20344 for further details

Targeted protein/peptide quantitation

  • General Information:
    • Targeted analysis allows for detection of peptides in samples in a more selective and sensitive experimental approach. Potential proteins and peptides can be identified in screening analysis, and relative amounts determined in subsequent analyses. Absolute quantitation of peptides can be performed by comparison with internal or external standards curves.
  • Accessible for training and student use, or fee-for-service work
  • Contact name: Dr. Muhammad Zenaidee - muhammad.zenaidee@mq.edu.au
  • NATA Accredited - refer to the Scope of Accreditation 20344 for further details

Post-translational modification (PTM) analysis

  • General Information:
    • APAF can support identification and relative quantitation of a wide range of PTMs including phosphorylation. PTMs can be enriched through specifically designed protocols to select for peptides with the desired PTM. Please contact us to discuss how we might be able to facilitate PTM analysis.
  • Accessible for training and student use, or fee-for-service work
  • Contact name: Dr. Muhammad Zenaidee - muhammad.zenaidee@mq.edu.au

Glycan analysis

  • General Information:
    • Glycans are polysaccharides that can be conjugated to proteins and affect protein structure and function. Glycans are cleaved either enzymatically or chemically, and the glycan profile of samples (glycan mass and composition) and relative abundance is determined by LC-MS. Glycoproteomics to determine site specific glycans can also be performed.
  • Accessible for fee-for-service work
  • Contact name: Dr. Muhammad Zenaidee - muhammad.zenaidee@mq.edu.au

Identification of protein interactions from co-immunoprecipitations and pull-downs

  • General Information:
    • Co-immunoprecipitations and pull-downs are robust methods for identifying protein interactions. APAF provides in-depth analysis of samples prepared via these techniques. Using sensitive mass spectrometry approaches, proteins in affinity enriched samples are identified and compared to those in controls. Comparisons are made via simple presence and absence, or in-depth relative quantification, depending on experimental needs.
  • Accessible for training and student use, or fee-for-service work
  • Contact name: Dr. Gene Hart-Smith – gene.hart-smith@mq.edu.au

Identification of protein interactions using crosslinking mass spectrometry

  • General Information:
    • Crosslinking reagents can enable interacting proteins, including transient interactions, to be covalently bound. APAF provides a range of techniques to identify interactions captured via crosslinking approaches. These include enrichment of crosslinked peptides, and sensitive mass spectrometric approaches for identifying crosslinked peptides, including those prepared using mass spectrometry cleavable crosslinkers.
  • Accessible for training and student use, or fee-for-service work
  • Contact name: Dr. Gene Hart-Smith – gene.hart-smith@mq.edu.au

In-solution measurements of biomolecular interactions and binding affinities

  • General Information:
    • Interactions between biomolecules, including proteins and polynucleotides, often require in-depth characterisation. APAF can provide these characterisations in-solution from purified samples using mass photometry. This can allow biomolecular interactions, including aggregation, to be confirmed, and enable sub-micromolar binding affinities and associated reaction kinetics to be determined.
  • Accessible for training and student use, or fee-for-service work
  • Contact name: Dr. Gene Hart-Smith – gene.hart-smith@mq.edu.au

Characterisations of intact proteins

  • General Information:
    • Many proteomics techniques characterise peptides derived from digested proteins. However characterisations of intact proteins can also be necessary. Intact protein measurements can, for example, confirm a protein’s sequence identity, characterise specific combinations of post-translational modifications, and measure protein-ligand complexes. APAF employs cutting- edge top-down and native mass spectrometry approaches, and mass photometric measurements, to enable such analyses.
  • Accessible for training and student use, or fee-for-service work
  • Contact name: Dr. Gene Hart-Smith – gene.hart-smith@mq.edu.au

Bioinformatics

  • General Information:
  • Accessible for training and student use, or fee-for-service work

Contact name:

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