Metabolomic Symposium
Friday, February 20th 2015
13:00 - 19:00
13:00
13:05
13:45
14:15
14:45
15:00
15:40
16:10
16:40
17:10
17:30
Opening remarks
David Heywood, Chairman
Omic and Targeted Proteomics and Metabolomics using UPLC/MS/MS: The Impact of UPLC Column
Dimensions
Arthur Mosely, Ph.D, Duke University
Plantomics, A case study on flavanoids
Maxim Maheux, Ph.D Candidate, Claude Paul Lafrance M.Sc. TransBIOTech
Untargeted 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
Break
Biomarker Discovery and Translational Research Leading to Clinical Utility
Christiane Auray-Blais, LL.M., Ph.D. Université de Sherbrooke
Application of mass spectrometry for development of plant-made influenza VLP vaccine.
Geneviève Mercier, MSc. Medicago
High throughput mass spectrometry coupled to machine learning for quality control assessment and
biomarkers discovery
Jacques Corbeil, Ph.D. Université Laval
ionKey/MS - beyond sample-limited applications
Geoff Gerhardt, PhD. Waters Core Research
Closing remarks
Cocktail
Centre de recherche du CHU de Québec
Fisher Auditorium
2705 Laurier Boulevard
Quebec city, Qc, G1V 4G2, Canada
Event questions:
Lynda Robitaille: Lynda.Robitaille.1@ulaval.ca
Patrice Lemire: patrice_lemire@waters.com
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.
1
Institut national de santé Publique du Québec, Québec, QC; Canada
Department of social and preventive medicine, Université Laval and Centre de Recherche du CHU de Québec,
Québec, QC. Canada
2
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-offlight mass spectrometry, and the development and validation of tandem 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. Nglycans 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 decision making 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.
Transbiotech is 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.