About the CDSS Bioanalytical Facility
The CDSS Bioanalytical Facility was initially established in Human Anatomy and Cell Biology in 1998, and was thus at the forefront of the new field of Proteomics. Following a successful bid led by Prof Kevin Park from the Department of Pharmacology and Therapeutics to become the MRC Centre for Drug Safety Science, the bioanalytical facility was incorporated into the Centre and was equipped with state-of-the-art instrumentation.
Initially focussing on drug metabolism and drug-protein interactions, the remit of the facility has expanded to include global discovery proteomics in health and disease, and absolute quantification of proteins and drugs/metabolites for biomarker validation
Equipment
The CDSS Bioanalytical Facility is equipped with top-of-the-range mass spectrometers and a wide selection of peripheral instrumentation for the pre-fractionation of samples prior to analysis.
The CDSS Bioanalytical Facility is equipped with a Sciex tripleTOF 6600 which is used for discovery proteomics pipelines (Figure 1)
Figure 1: Discovery proteomics pipeline
We have expertise in label-dependent (iTRAQ, ICAT, SILAC) and label-free (SWATH, DIA) quantitative approaches that allow a global view of protein expression. We are also able to quantify peptides of interest at low attomole (femtogram) levels of sensitivity using targeted multiple reaction monitoring on a Sciex hydrid linear ion trap-triple quadrupole mass spectrometer (QTRAP 6500, Figure 2). This approach is particularly useful when good quality antibodies are not available.
Figure 2
In addition, we have a further instrument (QTRAP 5500) dedicated to the analysis of small molecules, with targeted analyses allowing for sensitive and specific absolute quantification of drugs and their metabolites, and of endogenous analytes. We also have expertise in a range of separation technologies which are able to improve proteome coverage and/or sensitivity of target detection by simplification of complex biological matrices (Figure 3).
Figure 3: A range of separation technologies which are able to improve proteome coverage and/or sensitivity of target detection by simplification of complex biological matrices
Specialities
There are three particular strengths in the facility. One is the application of SWATH/DIA (data-independent) mass spectrometry to clinical proteomics. This approach provides a comprehensive catalogue of proteins present in a cell or tissue which is then used to identify and quantify thousands of proteins in individual human samples, allowing for patient stratification and the potential for identification of translatable biomarkers. Another is the characterisation of unconventional protein modifications. These may be as a result of direct drug or metabolite binding, but may also be a result of perturbed physiological processes leading to unexpected biochemical modifications of protein. The integration of biology, chemistry and mass spectrometry is a powerful approach in this area of research. Finally, the facility is committed to training the next generation of biological mass spectrometrists and has made significant contributions to the theses of over 40 PhD students in the last 10 years.
Back to: Faculty of Health and Life Sciences