Located on the top floor of the Peter Gilgan Centre for Research and Learning, SPARC BioCentre is a world-class facility operated by The Hospital for Sick Children. SPARC has been in continuous operation for over 27 years (albeit under different names), yet remains state-of-the-art thanks to strategic investments by SickKids and the Canada Foundation for Innovation (CFI). The SPARC Molecular Analysis (MA) division specializes in the investigation of proteins by mass spectrometry and amino acid analysis. With advanced instrumentation and knowledgeable and experienced staff, SPARC MA is well positioned to support innovative proteomics research.
Typical mass spectrometry (MS) experiments at SPARC MA range from resolution of simple samples, such as identifying protein gel bands or determining the accurate mass of a purified protein, to characterization of complex biological systems through peptide mapping or whole proteome profiling. Recent acquisition from Thermo Scientific of two Orbitrap Fusion Lumos Tribrid mass spectrometers equipped with advanced peptide fragmentation technology enables multiplexed relative quantitation of peptides using isobaric tags. SPARC MA instrumentation can also identify proteins that have undergone post-translational modifications, such as phosphorylation, which can impact the function or level of activity of a protein. Both of these untargeted approaches to proteomic analysis allow investigators to compare the proteomes of normal and diseased cells to increase their understanding of devastating diseases affecting Canadians such as cancer, heart failure, and neuropathies. Once specific proteins or modifications of interest have been identified, SPARC staff can help design targeted proteomics workflows, whereby a specific peptide in a complex mixture is identified and quantified across multiple samples. These studies are often performed on SPARC’s highly sensitive mass spectrometers, the TSQ Altis Triple Quadrupole or Q Exactive HF-X Hybrid Quadrupole-Orbitrap instruments from Thermo Scientific. However, clients do not need to worry about which particular MS instrument will be used for their proteomic research; SPARC staff are happy to discuss their experimental needs and budget, provide technical expertise, and advise on the best course of action. Once the experiment is completed, staff also analyze the data and ensure the client understands the results.
The amino acid analysis lab at SPARC routinely performs accurate analyses of protein or peptide sample concentration, composition, and purity. The lab services academic and industrial researchers, with high demand for physiological free amino acid analysis of biological samples and quantitation of unusual amino acids like selenomethionine. As with MS services, SPARC staff can work with clients to develop customized sample preparation techniques, methodologies, and results to suit their needs.
SPARC MA operates as a cost-recovery, fee-for-service facility, and is continually adding new services to keep up with research demands. For instance, SPARC recently began offering fractionation of complex protein samples by high-performance liquid chromatography for deeper proteome coverage, as well as enrichment of phosphorylated peptides. To learn more about SPARC and the many services offered, including Edman sequencing and extracellular flux (Seahorse) analysis, please visit the SPARC website. For general inquiries or to discuss your proteomics research needs, please contact SPARC’s manager, Leanne Wybenga-Groot at leanne.wybenga-groot@sickkids.ca.