Clinical samples are often scarce, making it difficult to analyse them with proteomics. For example, a typical proteomics experiment may require the equivalent of protein from 1 million cells. However, when dealing with clinical specimens we are often limited to samples of fewer than 10,000 cells. This project aims to develop the clinical proteomic workflow to enable comprehensive analysis of very scarce samples.
Key approaches include the refinement of protein extraction, separation, and analysis. We employ alternative protein extraction approaches (non-detergent) under ultra-clean conditions (HEPA-filtered environment). We can use multi-dimensional HPLC separation coupled with QTOF mass spectrometry for ultra-sensitive detection of proteins, peptides, and post-translational modifications. Importantly, these approaches are refined specifically for the sample of interest, whether it be a solid tumour/tissue, sorted cells, CSF, laser-capture dissected frozen sections, etc.
The ability to conduct comprehensive proteomic analyses on scarce samples is an important platform method for our lab and used broadly in our in-house and collaborative projects. If this approach is of interest to you please drop us a line.
This research project is available to Masters and PhD students to join as part of their thesis.
Please contact the Research Group Leader to discuss.
Dr Dalia Ponce Garcia, Research Officer
Associate Professor James Ziogas
Associate Professor Fred Hollande
Dr Tom Hennesey
University of Melbourne
- Mangum JE, Kon JC, Hubbard MJ. Proteomic analysis of dental tissue microsamples. Methods Mol Biol 2017; 1537: 461-479.
- Mangum JE, Crombie FA, Kilpatrick N, Manton DJ, Hubbard MJ. Surface integrity governs the proteome of hypomineralized enamel. J Dent Res 2010; 89(10): 1160-1165.
- Shnyder SD, Mangum JE, and Hubbard MJ. Triplex profiling of functionally distinct chaperones (ERp29/PDI/BiP) reveals marked heterogeneity of the endoplasmic reticulum proteome in cancer. J Proteome Res 2008; 7(8): 3364-3372.
- Mangum JE, Veith PD, Reynolds EC, Hubbard MJ. Towards second-generation proteome analysis of murine enamel-forming cells. Eur J Oral Sci 2006; 114(Suppl 1): 259-265.
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