Metabolomic Symposium

Metabolomic Symposium

Friday, February 20th 2015

13:00 - 19:00

13:00 Opening remarks

David Heywood, Chairman

13:05Omic and Targeted Proteomics and Metabolomics using UPLC/MS/MS: The Impact of UPLC Column Dimensions

Arthur Mosely, Ph.D, Duke University

13:45 Plantomics, A case study on flavanoids

Maxim Maheux, Ph.D Candidate, Claude Paul Lafrance M.Sc.TransBIOTech

14:15Untargeted metabolomics combined to quantification of amino acids and acylcarnitines

Cynthia Roy, Ph.D. Candidate, Pierre-Yves Tremblay M.Sc. INSPQ, Pierre Ayotte, Ph.D. Université Laval

14:45Break

15:00Biomarker Discovery and Translational Research Leading to Clinical Utility

Christiane Auray-Blais, LL.M., Ph.D. Université de Sherbrooke

15:40Application of mass spectrometry for development of plant-made influenza VLP vaccine.

Geneviève Mercier, MSc. Medicago

16:10High throughput mass spectrometry coupled to machine learning for quality control assessment and biomarkers discovery

Jacques Corbeil, Ph.D. Université Laval

16:40ionKey/MS - beyond sample-limited applications

Geoff Gerhardt, PhD. Waters Core Research

17:10Closing remarks

17:30Cocktail

Centre de recherche du CHU de Québec

Fisher Auditorium

2705 Laurier Boulevard

Quebec city, Qc, G1V 4G2, Canada

Event questions:

Lynda Robitaille:

Patrice Lemire:

The Speakers

Arthur Mosely, Duke University, Durham, NC, USA

Maxim Maheux, Claude-Paul Lafrance, Transbiotech Qc, Canada

The flavonoids constitute a vast class of endogenous plant metabolites. The interest for these compounds stems from their health protective properties, which have been confirmed in numerous studies. The flavonoids are divided in six families, depending on the nature of the substituents and double bonds in their central oxane ring. Owing to the presence of hydroxy and methoxy groups on the two aromatic rings, the latter frequently glycosylated, several thousands of flavonoids have been found in nature. Since the identification by comparison with standards is therefore impossible, some other analytical strategies have to be designed. We will present two of these, the first being a UPLC separation coupled with constant neutral loss measurements using triple quadrupole instruments, in order to determine the unit mass of the aglycones and the type of substitution with sugar moieties. The second involves the use of exact mass measurements and ion mobility experiments.

Cynthia Roy1,, Pierre-Yves Tremblay1 and Pierre Ayotte1,2.

1Institut national de santé Publique du Québec, Québec, QC; Canada

2 Department of social and preventive medicine, Université Laval and Centre de Recherche du CHU de Québec, Québec, QC. Canada

Plasma levels of branched-chain aromatic amino acids, aromatic amino acids and acylcarnitines have been linked to the future risk of type 2 diabetes. In order to measure these biomarkers while mining for new ones, we have developed a method that combines the quantification of eight amino acids and eight acylcarnitines together with untargeted metabolomics. The hybrid method was developed on a Waters UPLC-ESI-QTOF platform. Targeted metabolites are quantified using Targetlynx™ with a 10 points calibration curve. A complete validation was performed for all targeted metabolites with regard to matrix effect, recovery, matrix effect on recovery, limits of detection and quantification, linearity, inter and intra-essay variability and short-term stability. Untargeted metabolomics processing and metabolite identification is performed using MZmine 2.0 and our in-house library. The processed data is then exported to MetaboAnalyst 3.0 for statistical analyses. Further identification of unknown potential biomarkers can be achieved using online databases, namely METLIN, MassBank and HMDB.

Christiane Auray-Blais, Université de Sherbrooke, Qc, Canada

Lysosomal storage disorders (LSDs) are a group of nearly 50 disorders displaying considerable clinical and biochemical heterogeneity. While the extent and severity of LSDs depend on the type and amount of accumulated substrate, almost all disorders are progressive. The overall objective of our research studies is to improve the management of LSD patients by analyzing biomarkers that reliably reflect disease progression and severity, and thereby facilitate the evaluation of new therapies by improved monitoring of the response to treatment. Results from the discovery of novel biomarkers using time-of-flight mass spectrometry, and the development and validation oftandem mass spectrometry methodologies for Fabry disease patients will be presented.

Geneviève Mercier, Medicago, Qc,Canada

Medicago has developed an influenza VLP (Virus-Like Particle) vaccine, using a novel manufacturing platform based on transient expression of hemagglutinin (HA) in Nicotiana benthamiana. VLP structure is composed of a lipid envelop with spikes of HA protein and is very similar to influenza virus when analyzed by electron microscopy. The protein content of the VLP vaccine is mainly HA (>90% purity). Proteins impurities can either come from the host cell or the manufacturing process. Analysis by LC-MSE along with data processing with Progenesis QI is used to identify those proteins. Hemagglutinin is a glycoprotein, with five to thirteen potential sites of N-glycosylation depending on the influenza strain. N-glycans are identified by LC-MSE using Biopharmalynx for data processing.

Jacques Corbeil, Université Laval, Qc, Canada. Canada Research Chair in Medical Genomics

Our ultimate goal is to improve procedures for quality control and discover disease biomarkers from blood product metabolomic analyses. Our approach will assist in diagnostic and prognostic assessments of blood transfusion adverse effects, chronic inflammatory condition such as Systemic Lupus Erythematosus and infectious diseases primarily HIV/AIDS by employing high throughput mass spectrometry and machine learning algorithms. Our software solution coupled to detailed protocols for sample preparation could be adapted to most complex biological products and assist in classifying specific states in health and disease.

Geoff Gerhardt, Waters Core Research, Milford, MA, USA

Designed for the most demanding quantitative applications, Waters mass spectrometers provide unprecedented levels of sensitivity and the ultimate in quantitative performance. ionKey/MS provides the same 2.1-mm I.D. UPLC chromatographic performance you know in a 150-µm I.D. channel packed with sub-2-µm particles, providing tremendous improvements in sensitivity. The iKey™ Separation Device is one of the enabling technologies behind the ionKey/MS™ System. The innovative iKey eliminates the need for traditional fittings and greatly simplifies the microflow-LC user experience. Available with an internal diameter of 150 µm and in 50 mm and 100 mm lengths, the iKey brings microflow LC into the mainstream of chromatographic laboratories.

Sponsors

The CRCHU de Québec is affiliated to Université Laval and proudly positions itself among the most important health research institutes in Canada. About 2700 highly qualified personnel work at the CRCHU de Québec. This includes over 500 researchers. More than 900 students/trainees at the master’s, Ph.D. postdoctoral and fellow levels attend the CRCHU de Québec, where they are receiving a world-class multidisciplinary training.
The CRCHU de Québec is the largest francophone research centre in North America and ranks among the ten most important research centres in Canada. Owing to the excellence of its research teams and its infrastructures equipped with state-of-the-art technology, it benefits from an enviable reputation both at the national and international levels.

The Institut national de santé publique du Québec (INSPQ) was created in 1998 following the adoption of its act of incorporation (RSQ, chapter I-13.1.1). Its experts come from fields as distinct as applied science, health sciences, social sciences, and humanities and work with the health and social services network to develop and pool public health knowledge and skills. The institute is an expertise and reference centre. As a broker of expertise, it contributes to the definition and understanding of public health issues, as well as to the identification of interventions with a positive impact on the determinants of population health deemed appropriate based on the contexts relevant to the decisionmaking process. It produces and shares scientifically based knowledge, through a range of quality products and services. It ensures the dissemination of this knowledge in a timely manner using the appropriate wording and a suitable format to effectively support decision makers, health care workers, and partners in their endeavours pertaining to population health determinants.

Transbiotechis a technology transfer college centre (CCTT) founded in 1999. It is affiliated with the Cégep [general and vocational college] Lévis-Lauzon, and recognized by the government of Quebec. Specialized in biotechnology, we are leading the way in innovation and support for the businesses of Quebec. Since 2002 and thanks to contributions from the government of Quebec and the Canada Foundation for Innovation, TransBIOTech has built a high-level research complex that meets the standards established by Health Canada for containment level 2 facilities. The organization is a member of the Trans-Tech Network, a network of CCTTs.

Waters creates business advantages for laboratory-dependent organizations by delivering practical and sustainable scientific innovation to enable significant advancement in healthcare delivery, environmental management, food safety, and water quality. Waters Division products are used by pharmaceutical, life science, biochemical, industrial, academic and government organizations working in research and development, quality assurance and other laboratory applications. Waters UPLC, HPLC and MS instruments and consumables are used in a broad range of industries to measure the chemical, physical and biological composition of materials. Waters Division is also headquartered in Milford, Massachusetts.