Isotopically-labelled TrEnDi: new technology to increase the sensitivity and selectivity of Mass Spectrometry-based lipid analysis of complex biological samples
Trimethylation enhancement using diazomethane (TrEnDi) is a chemical derivatization strategy that results in the complete and concomitant methylation of phosphate moieties, carboxylic acids and primary amines, rendering phosphatidylethanolamine (PE), phosphatidylcholine (PC), and phosphatidylserine (PS) glycerophospholipids, as well as the sphingolipid sphingomyelin, permanently positively charged. Modified lipids demonstrate enhanced sensitivity via mass spectrometry (MS) analysis, particularly in MS2 experiments where ion fragmentation is consolidated to only one or two channels. One challenge initially encountered by this technique was the conversion of PC and PE molecules of identical chain length to identical (isobaric) lipids. To overcome this, a novel method to synthesize 13C-labelled diazomethane was developed. This presentation will demonstrate the sensitivity gains observed using isotopically-labelled TrEnDi on complex lipid mixtures as well as highlight some of recent modifications that have improved the performance of this derivatization chemistry.
Biosketch: Dr. Jeff Smith is Associate Professor in the Department of Chemistry and the Institute of Biochemistry at Carleton University. He received his BSc from Trent University in 2000 and earned his PhD at York University in 2005. After three years at the Ottawa Institute of Systems Biology at the University of Ottawa, he joined Carleton in 2008, and is currently the Director of the Carleton Mass Spectrometry Centre. His research focusses on the use of mass spectrometry to investigate the biomolecular mechanisms of cellular life.
Accelerated Data Workflows for Biomarker Discovery in Metabolomics: Clinical Applications in Newborn Screening and Cystic Fibrosis Research"
Sponsored by Agilent Technologies
Philip Britz-McKibbin, Professor and Cystic Fibrosis Canada Researcher, Dep. Chemistry & Chemical Biology, McMaster UniversityAbstract: Comprehensive metabolite profiling (i.e., metabolomics) offers a holistic approach for understanding the phenotype of an organism on a molecular level that is closely associated with clinical outcomes. However, low sample throughput, complicated data processing and unknown compound identification remain major bottlenecks to biomarker discovery in clinical-based metabolomics research. An introduction to multisegment injection-capillary electrophoresis-mass spectrometry (MSI-CE-MS) will first be presented as a multiplexed separation platform that takes advantage of customized serial injections to enhance sample throughput and data fidelity with quality assurance. Various sample injection formats can be designed to encode MS information temporally when using signal pattern recognition for unambiguous biomarker identification when analyzing volume-restricted biological samples. Structural elucidation of unknown metabolites can be deduced with greater confidence when using high resolution tandem MS experiments in conjunction with CE-based mobility theory. Recent progress towards characterization of the sweat metabolome of screen-positive cystic fibrosis infants, as well as identification of new markers of CF in asymptomatic neonates from retrospective dried blood spot specimens will be discussed in support of universal newborn screening programs. MSI-CE-MS offers a transformative approach for expanding the productivity of MS while offering an accelerated data workflow for biomarker discovery in metabolomics.
Speaker Bio: Philip Britz-McKibbin is a Professor at the Department of Chemistry and Chemical Biology and Cystic Fibrosis Canada Researcher based at McMaster University. Prior to starting his current position, he completed his doctorate research with David D.Y. Chen at the University of British Columbia (2000) followed by a visiting lecturer position at the University of North Carolina at Greensboro in USA (2001) and a post-doctoral fellowship with Shigeru Terabe at Himeji Institute of Technology in Japan (2003). His research interests in bio-analytical chemistry and metabolomics include the design of new analytical strategies to quantify and identify metabolites in biological samples using innovative separation, mass spectrometric and bioinformatic tools. Philip’s laboratory aims to discover new biochemical markers that support early detection and treatment of human diseases relevant to population/occupational health and pediatric/perinatal medicine with emphasis on inherited metabolic disorders and the developmental origins of health and disease. His research has been funded by NSERC, CFI, CIHR, Cystic Fibrosis Foundation of Canada, the Ontario Genomics Institute, and the Ontario Ministry of Labour. He has presented over 100 invited talks and published more than 70 peer-reviewed articles. Dr. Britz-McKibbin is the recipient of several prestigious awards, including the American Chemical Society – Young Investigator Award in Separation Science (2010), Japan Society for Promotion of Science (2009), Petro-Canada Young Investigator Award (2007) and Premier’s Research Excellence Award (2004). Philip is a recent Evaluation Group Member in Chemistry for NSERC Discovery Grant program, an editorial board member in Chemical Biology of Scientific Reports, a Founding Advisory Board Member of the US Chapter of the Metabolomics Society and an affiliate member of the Metabolomics Innovation Centre – Canada’s national metabolomics facility.
In-depth analysis of LC-MS/MS data
Invited Speaker for 2019 2nd Floor Research Day
March 19th, 2019; 10:15 am; HSB GB03
Baozhen (Paul) Shan, Bioinformatics Solutions Inc., ON, Canada
Abstract: A brief introduction to mass spectrometry for health sciences research and an an introduction to new methods for data analysis for proteomics.
Speaker Bio: Dr. Baozhen Shan is the CEO of Bioinformatics Solutions Inc. (BSI), one of the leading proteomics software and service platform providing companies. He began working at BSI in 2008. He obtained his first PhD in Chemistry in 1997 from Nanjing University, and his second in Computer Science in 2009 from the University of Western Ontario.
As a computational chemist, he has spent more than 15 years in research and development related to data mining and statistical analysis of mass spectrometry-based proteomics (including protein/peptide identification and quantification, antibody characterization, etc.). As a result he has developed a broad set of scientific and algorithmic skills of modelling and understanding molecular biology through statistical analysis.
Workshop -Using PEAKS software on proteomics data analysis
9:00 AM-4:30 PM, March 20, 2019
E1430 Health Sciences (Library Board Room)
Baozhen (Paul) Shan, CEO, Bioinformatics Solutions Inc., ON, Canada
- Workflow of LC-MS/MS (9 am – 10 am)
- DDA vs DIA
- Protein/Peptide Identification Solutions (10:10 am-12:10pm)
- Database search and de novosequencing
- Identification of PTMs and sequence variants
- In-depth analysis with Ion Mobility LC-MS
- Protein/Peptide Quantitation Solutions (1:30pm-4:30pm)
- Label-Free Quantification
- Labelled Quantification (ex. SILAC, iTRAQ, TMT, MS3)
- Post-Translational Modification (PTM) Quantification
Note: Please bring your own laptop for hands-on practice. Slides and demo software will be provided.