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Magnetic Bead Mixer for Mini Nucleic Acid Purification
July 2011 Volume 15, Number 4
Optimizing Liquid Chromatography
Sample and Mobile Phase Preparation
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e n v i r onm e n t
R&D News.......................... 1 Appointments..................... 6 Pharma Notes..................... 7 New Products................... 14 Calendar........................... 17 Career Spotlight............... 18
find out more about the promise of stem cell research, and add your support to the call for an accelerated approach to eradicating the major disease of our time. • Follow the discoveries. Receive updates on the work being done in the areas of heart, blood, diabetes, neurology and lung and the progress being made toward cures. • Direct the research. Channel your support directly to the areas of research that matter the most to your personally. Representatives of the McEwen research community were joined for the Stem Cell City announcement by representatives of the Ontario Government, the University Health Network (UHN), the international stem cell community, and patient advocates to help launch this online initiative.
Dr. Gordon Keller
Stem Cell City Opens UHN’s McEwen Centre for Regenerative Medicine announced the creation of Stem Cell City, a virtual community designed to inform the
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public about stem cell research and allow them to articulate their support of using regenerative medicine to fight disease.
According to Dr. Gordon Keller, director of the McEwen Centre for Regenerative Medicine, Stem Cell City is an online web portal designed to teach the public about the body’s tiny, tissue-generating powerhouses and to inform them of the latest research aimed at defeating the most devastating diseases afflicting humans — from heart and lung disease to Alzheimer’s and diabetes. The site includes virtual tours of the centre’s labs, allows visitors to key into research areas of personal interest as well as provides options to donate to research. Other notable features of the site include: • Join the community. Explore the world of regenerative medicine,
The “city” was opened June 15, 2011 by the Centre’s co-cofounders Rob and Cheryl McEwen. They were joined by Dr. Keller, Ontario Premier Dalton McGuinty, Minister of Citizenship and Education Dr. Eric Hoskins, UHN CEO and president Dr. Robert Bell, California Institute for Regenerative Medicine (CIRM) chairman Robert Klein and patient advocate Lisa Ray. For more information or to join Stem Cell City go to www. joinstemcellcity.com.
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July 2011 Laboratory Focus www.bioscienceworld.ca
news Team from NRC recognized for developing neurosurgical simulator The National Research Council of Canada (NRC) is recognizing 52 employees for exceptional success during its Outstanding Achievement Awards. The NeuroTouch team leads were awarded the “NRC Breakthrough of the Year Award” for their creation of a neurosurgical
simulator called NeuroTouch. This state-of-the-art technology allows neurosurgeons to rehearse complex surgeries on simulated brain tumours that are specifically tailored to their patients, reducing the risk of complications and speeding up recovery. Their work is part
of the Genomics and Health Initiative, a cross-country multidisciplinary collaboration. As a result of this initiative, seven virtual reality training centres will be established across the country, four of which already exist in Halifax, Montréal, Ottawa and Toronto.
The NeuroTouch team leads have helped Canada remain a world leader in next-generation virtual medical simulation. The NeuroTouch technology is laying the groundwork for sharing know-how and advanced made-in-Canada solutions with the world.
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Scientists map path to generate renewable antibodies
Dr. Tony Pawson, Senior Investigator at the Samuel Lunenfeld Research Institute of Mount Sinai Hospital
In the first study of its kind to date, an international group of scientists including Drs. Tony Pawson and Karen Colwill at Mount Sinai Hospital and Drs. Dev Sidhu and Aled Edwards at the University of Toronto have highlighted the power of renewable antibody technologies to create precise reagents for use in biomedical research. This methodology will help accelerate scientists’ understanding of all the proteins coded in the human genome. The study was published online in the biomedical journal Nature Methods. Antibodies are essential tools in biomedicine, used both for discovery and therapy. However, many commercially available antibodies for research are of inadequate quality and supply, which hinders scientists from iden-
tifying new causes of and potential therapies for diseases including cancer. The problem is compounded because the genomic revolution has dramatically increased the demand for renewable, highquality affinity reagents to the many newly discovered and uncharacterized proteins. “The use of renewable antibodies hasn’t been fully exploited,” said Dr Tony Pawson, distinguished investigator at the Samuel Lunenfeld Research Institute of Mount Sinai Hospital and study coauthor. “Our study shows we can rapidly make reagents that are highly specific to a large number of proteins, which will facilitate a more precise examination of the proteins and pathways involved in various complex illnesses.” “The potential of the human
genome project has not yet been fully realized because we haven’t had the full set of necessary tools,” echoed Dr Sidhu, principal investigator at the University of Toronto’s Donnelly Centre for Cellular & Biomolecular Research, and director of the Toronto Recombinant Antibody Centre (TRAC). “The team has now generated a roadmap to dramatically increase the number and quality of these tools, and this is an important advance.” Participating institutions include the University of Toronto, University of Chicago (USA), University of Cambridge (Britain), Karolinska Institutet (Sweden), Technische Universität Braunschweig (Germany), Monash University (Australia) and the Royal Institute of Technology (Sweden). The study received support from the Canadian Institutes for Health Research, Canada Foundation for Innovation, Genome Canada through the Ontario Genomics Institute, GlaxoSmithKline, the Karolinska Institutet, the Knut and Alice Wallenberg Foundation, the Ontario Innovation Trust, the Ontario Ministry for Research and Innovation, Merck & Co, the Novartis Research Foundation, the Swedish Agency for Innovation Systems, the Swedish Foundation for Strategic Research and the Wellcome Trust.
Miraculins receives $100,000 Grant through technology commercialization program Medical diagnostic company Miraculins Inc. will receive $100,000 in grant funding from the Manitoba Technology Commercialization Program to support the commercialization of its PreVu non-invasive cholesterol test. PreVu evaluates the additional risk a person may have for coronary artery disease (CAD) by simply and painlessly testing their skin cholesterol level. High levels of skin cholesterol have been shown to be correlated to CAD as measured by stress test, angiography, coronary calcium, carotid intima media thickness, inflammatory
markers of vascular disease, previous heart attack incidents and Framingham risk score. The Technology Commercialization Program is a Province of Manitoba initiative which assists Manitoba-based companies in the commercialization of technologies. The financial assistance program provides funding to cover up to 50 per cent of eligible costs including legal fees, patenting, marketing studies, literature development, product certification, prototyping, process validation and technical consulting costs. The maximum cost-sharing available under the program is $100,000.
Call for papers for Pittcon 2012
OCCC The program committee for Pittcon 2012 is inviting the scientific community to submit abstracts for contributed oral or poster presentations. Joseph J Grabowski, PhD,
program committee chairman, says his committee is seeking a diverse, yet comprehensive technical program covering all areas of analytical science and applied spectroscopy. A partial list of symposium topics will include: Instrumental analysis; chromatography, molecular and atomic spectroscopy, portable/hand-held instruments, electrochemistry, mass spectrometry, and lab-on-chip technology, and novel application of measurement science;
nanotechnology, biomedicine, neurochemistry, genomics, proteomics, metabolomics, metabonomics, forensics, informatics, pharmaceutical, environmental, polymer, and food science. The deadline for submissions is August 12, 2011. Authors will be notified in October 2011, by email only, as to the status of their abstracts. Pittcon 2012 takes place on March 11-15 at the Orange County Convention Center in Orlando, FL.
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July 2011 Laboratory Focus www.bioscienceworld.ca
news
Theralase and George Brown College join in Governmentfunded research to combat food contamination Theralase Technologies Inc. is partnering with George Brown College’s Centre for Hospitality and Culinary Arts to conduct applied research in the use of patented photodynamic compounds (PDCs) activated by its proprietary laser technology to destroy microbial pathogens associated with food contamination. This food safety approach, believed to be the first of its kind, is supported by the National Sciences and Engineering Research Council of Canada (NSERC). PDCs are light sensitive molecules that have the ability to attach themselves to specific cell types,
in this case food pathogens such as bacteria, and are able to destroy these cells upon light activation. Theralase Technologies Inc, founded in Toronto in 1995, designs, develops and manufactures patented, superpulsed laser technology utilized in biostimulation and biodestruction applications. “In the wake of many outbreaks of consumer food contaminations associated with microbial pathogens such as Listeria monocytogenes (Listeria) and Escherichia coli (E. coli) and their various strains, Theralase initiated the study to determine the feasibility of
using our patented PDCs for sanitation in the food service, food manufacturing and medical industries,” said Roger Dumoulin - White, president & CEO of Theralase Inc. “Given their history and pedigree in culinary arts and additional focus on food applied research, the George Brown College Centre for Hospitality and Culinary Arts is a perfect partner for applied research in new and innovative sanitation protocols and techniques using the company’s patented lasers and photodynamic compounds.” The 14-week food safety feasibility study will determine the specific areas
in food processing facilities where pathogen contamination is not being adequately addressed by current processes and procedures. The results of the study are expected in September 2011 and will be followed by further lab testing and a pilot project at a commercial food processing facility.
U of S Partners with Federal Agencies on unique study of Vitamin D insufficiency Dr. Frédéric Charron, and Luisa Izzi.. Photos: IRCM
Breakthrough at the IRCM could help understand certain cancers A scientific breakthrough by researchers at the Institut de recherches cliniques de Montréal (IRCM) will be published in Developmental Cell, a scientific journal of the Cell Press group. Led by Dr. Frédéric Charron, the team of scientists discovered a new requirement for the proper functioning of the Sonic Hedgehog protein. Sonic Hedgehog belongs to a family of proteins that gives cells the information needed for the embryo to develop properly. It plays a critical role in the development of many of the body’s organs, such as the central nervous system. Malfunctions of these proteins are associated with many diseases including cancer, which is the leading cause of death in Canada. “On one hand, certain molecules travel through our organs (in this case, Sonic Hedgehog) and transmit signals to cells with information on how they should function,” explains Luisa Izzi, postdoctoral fellow in Dr. Charron’s laboratory and co-first author of the article. “On the other hand, our cells have receptors to receive these signals. The receptors then instruct the cell’s DNA as to which genes to turn on or off in
order to perform its function.” The team studied the interactions between the Sonic Hedgehog molecule and the recently-identified receptors Boc, Cdon and Gas1, all found on the surface of cells. “Our research showed, unexpectedly, that these receptors are essential for the transmission of the hedgehog molecule’s signal,” adds Martin Lévesque, an alumnus from Dr. Charron’s research unit and co-first author of the article. “Disrupting the transmission of the Sonic Hedgehog signal can lead to diseases,” says Dr. Charron, director of the IRCM’s Molecular Biology of Neural Development research unit. “A better knowledge of the receptors Boc, Cdon and Gas1 might in turn help our understanding of pathologies associated with defective Sonic Hedgehog signalling. Our results could also lead to new avenues for the treatment of certain diseases such as cancer.” Research carried out in Dr. Charron’s laboratory was funded by the Canadian Cancer Society, the Fonds de recherche en santé du Québec (FRSQ), and the Canadian Institutes of Health Research (CIHR).
One in four Canadians aged six to 79 has vitamin D blood levels below internationally recommended levels, a first-in-Canada study led by the University of Saskatchewan, in partnership with Statistics Canada and the Public Health Agency of Canada, has found. When individuals taking vitamin D supplements were excluded from the data, evidence of low vitamin D blood levels was even greater. In fact, during the winter months, more than one-third of Canadians not using supplements did not meet the level associated with the recommended daily allowance, the study found. “We have shown the advantage that supplement use and sun exposure can have to help Canadians maintain their vitamin D status,” said U of S nutrition professor Susan Whiting. Low vitamin D can increase the risk of bone-related problems such as rickets, osteomalacia (softening of the bones), and osteoporosis. Low vitamin D may also compromise the immune and inflammatory systems. The new study, published in the American Journal of Clinical Nutrition, suggests that current food choices alone are insufficient to maintain recommended vitamin D levels for many Canadians, especially in winter. The study is the first in Canada to examine vitamin D intake in comparison to levels outlined
in the U.S. Institute of Medicine’s 2011 Dietary Reference Intake (DRI) report which sets out the estimated average daily vitamin D requirements for individuals. The Canadian government has accepted that report as the basis for new recommended values for vitamin D and calcium levels considered adequate for specific age groups, as well as safe upper levels. The research was conducted by Whiting and Hassan Vatanparast of the U of S College of Pharmacy and Nutrition in collaboration with Linda Greene-Finestone of Public Health Agency of Canada and Kellie Langlois of Statistics Canada.
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July 2011 Laboratory Focus www.bioscienceworld.ca
APPOiNtMeNtS
Borden Ladner Gervais LLP (BLG) announces that Jay Lefton has joined the firm as a partner in its
Toronto office. Lefton has more than 25 years of experience advising public and private companies on M&A, corporate and securities law, with a particular focus on technology and life sciences. In 2011, Jay was selected by his peers for inclusion in The Best Lawyers in Canada® and recognized in the International Who’s Who of Mergers & Acquisitions Lawyers and the Lexpert®/American Lawyer Guide to the Leading 500 Lawyers
in Canada. Lefton currently sits on many boards including the Cell Therapy Advisory Board of Toronto’s Princess Margaret Hospital, the Ontario Genomics Institute, ThinkFirst Foundation of Canada, the Ontario Neurotrauma Foundation, Pathways to Education, Canadian Friends of Hebrew University and the Canadian Committee of Tel Aviv Foundation. He is also on the Board of Governors and the Research and Commercialization Committee of Toronto’s Mount Sinai Hospital. He has been a member of the Ontario Securities Commission’s Securities Advisory Committee assisting on various matters, including legislative and policy initiatives. He has also been an adjunct faculty member at Osgoode Hall Law School and has also instructed at the Rotman School of Management at the University of Toronto. Novadaq Technologies Inc. has appointed Dr. David C. Martin as vice president, Business Development and Investor Relations. Prior to joining Novadaq, Dr. Martin worked as a healthcare analyst and investment banker with Dundee Securities and Yorkton Securities. Before that, he worked in sales and marketing for clinical diagnostics and biotech-
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nology instrumentation companies, including Boehringer Mannheim and InterScience Inc. Dr. Martin has a Ph.D. (Biochemistry) from the University of Western Ontario, a M.Sc. (Biology) from the University of Toronto, and an MBA from the Richard Ivey School of Business. Patheon announces the appointment of Michael E. Lytton as executive vice president, Corporate Development and Strategy and General Counsel. Lytton was most recently with Biogen Idec, Inc., where he held the position of executive vice president, Corporate and Business Development. Lytton’s 28-year career includes extensive experience in the pharmaceutical and biotech industry. Prior to joining Biogen Idec, Lytton was a general partner with Oxford Bioscience Partners, a venture capital firm investing in therapeutic, diagnostic, and life science tool companies. He has also practiced law for 17 years and specialized in representing biomedical companies; and is a past partner and member of the executive committee of the law firm Edwards Angell Palmer & Dodge as well as partner of the law firm Wilmer Hale. Currently he is chairman of the Board of Santhera Pharmaceuticals AG. He holds an undergraduate degree from Princeton University, a Master of Science degree in Epidemiology from the London School of Hygiene and Tropical Medicine at the University of London, U.K. where he was a Fulbright Scholar, and a Juris Doctor from Harvard Law School. Cytochroma announces the appointments of Dr. Cynthia Lavoie and Osamu Otani to its Board of directors. Dr. Lavoie replaced Dr. Luc Marengère as VG Partners’ representative, and Mr. Otani replaced Dr. Seiichi Kiso as Mitsubishi Tanabe Pharma Corporation’s representative on the company’s Board. The number of Board members remains unchanged at six members. Dr. Lavoie is a Partner with VG Partners, part of VenGrowth Private Equity Partners Inc., and co-heads the management of the VenGrowth Advanced Life Sciences Fund Inc. She brings over 16 years of experience in research and industry and works closely with company management lending her technical and financial expertise to operations as well as financing, merger and exit transactions. Currently she sits on the boards of Axela, Interface Biologics, Trillium Therapeutics and Zelos
Therapeutics. Her prior experience includes positions as Investment Manager with the Ontario Genomics Institute where she initiated a seed financing program and as Marketing Strategy consultant with a medical device and therapeutics company. Mr. Otani joined Tanabe Seiyaku Company Ltd. in 1983, and has served in managerial positions across International Development and project management, including general manager, Tanabe Seiyaku London Office, and managing director, Mitsubishi Pharma Research & Development (Beijing). He has been president, Mitsubishi Tanabe Pharma Development America, Inc. since July 2010. Asmacure Ltee, a clinical-stage biopharmaceutical company focused on the development of nicotinic receptor agonists for the treatment of asthma and other inflammatory diseases, announces that Martin Driscoll has been appointed as chief executive officer. Driscoll will lead the company as it progresses its lead compound further in clinical development. Driscoll has more than 30 years experience in the pharmaceutical industry, including senior roles in general management, commercial operations and business development for both privately held and publicly traded life science companies. He led the commercialization of several successful therapeutic products, the direct negotiation of multiple transactions, licensing and M&A and the successful submission of multiple major product regulatory filings. Prior to being elected CEO of Asmacure, Mr. Driscoll was CEO and a director of Javelin Pharmaceuticals, Inc., a publicly traded company focused on the development of acute care pain products, from March 2008 until July 2010, at which point he led the successful merger of the company with Hospira, Inc. Prior to his role with Javelin, Mr. Driscoll held senior managerial roles at Schering-Plough Corporation, ViroPharma, Inc. and Reliant Pharmaceuticals. During his tenure at Schering-Plough, he also spent a significant amount of time as general manager of Key Pharmaceuticals, the Schering unit responsible for all respiratory products marketed in the U.S. He holds a B.Sc. in Communications from the University of Texas at Austin. He has also been recently appointed as a member of the board of the Epilepsy Foundation of New Jersey and served on the Advisory Board for the American Lung Association.
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July 2011
Pharma Notes Sanofi-aventis (Laval, QC) announces that Health Canada has granted market authorization for Jevtana™ (cabazitaxel) injection in combination with prednisone for the treatment of patients with metastatic castration resistant (hormone refractory) prostate cancer (mCRPC), previously treated with a docetaxel-containing treatment regimen. mCRPC is an advanced form of prostate cancer in which the cancer has continued to grow despite the suppression of male hormones, called androgens, which often fuel the growth of prostate cancer cells. Jevtana™ has been shown to work by causing the intracellular microtubules, which help separate chromosomes into daughter cells during cell division, to stabilize so that the cells can no longer grow or divide. Jevtana™ was approved based on results from the Phase 3 TROPIC (Treatment of Metastatic HormoneRefractory Prostate Cancer Previously Treated with Taxotere-Containing Regimen) clinical study involving 755 patients with mCRPC previously treated with a docetaxel-containing treatment regimen, the results of which were published in the Lancet. Immunovaccine Inc. (Halifax, NS) has completed analysis of immune responses from patients enrolled in its Phase 1 clinical trial assessing the safety and tolerability of DPX-0907, a therapeutic cancer vaccine. The study was designed to evaluate the safety and immunogenicity of DPX-0907 in patients with advanced stage breast, ovarian or prostate cancer. DPX-0907 combines seven peptide antigens plus an adjuvant with Immunovaccine’s DepoVax™ delivery platform. The Phase 1 trial was conducted at five centres in the U.S. In this open-label, dose-escalating trial, patients received three injections (0.25 mL or 1 mL doses) of the active immune therapy DPX-0907, three weeks apart. Safety was assessed in 11 patients in the 0.25 mL dose group and 11 patients in the 1 mL dose group. The immunogenicity results
were based on an analysis of nine evaluable patients in the 0.25 mL dose group and nine evaluable patients in the 1 mL dose group. The Phase 1 trial met the primary objective of safety with overall results demonstrating
that DPX-0907 is generally well tolerated by all patients and is considered safe at both dose levels. There were no vaccine related serious adverse experiences reported to date. The secondary objective was to assess
whether administration of DPX-0907 could generate an immune response specific to the seven cancer antigens. Immunovaccine performed a detailed analysis of patients’ blood samples that showed cell mediated immunity
7
(CMI) to vaccine targets in all 3 breast cancer patients, 5 of 6 ovarian cancer patients and 3 of 9 prostate cancer patients. Both dose levels produced a targeted immune response in vaccinated patients.
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July 2011 Laboratory Focus www.bioscienceworld.ca
Feature
B y S i k an d e r G i ll , Raj w an t G i ll , H e n r y Yan g , an d Don g L i an g
Table 1
Validation of
Automated Magnetic Bead Mixer for VERSA Mini Nucleic Acid Purification/PCR Setup Workstation
Applications of magnetic beads
Separation and purification of nucleic acids1 RNA purification 2 Peptide aMpt-Mediated Capture PCR for Detection of Mycobacterium avium in Milk Samples3 SureSelect Target Enrichment for Illumina Paired-End Sequencing Library4 Target Enrichment for Massively Parallel Sequencing and SNP4 SPRI DNA fragment size selection5 PCR sample cleanup6 Novel purification for 6xHis-tagged7
Immobilization of l-lactate dehydrogenase on magnetic nanoclusters for synthesis o Gene Silencing Mediated by Magnetic Lipospheres Tagged with 15 pharmaceutical compounds 8 siRNA Functional Selection of Vaccine Candidate Peptides from Staphylococcus aureus
9 Bio-affinity magneticExpression reactor forLibraries peptide In digestion Whole-Genome Vitro16
17
Magnetic isolation circulating tumor Selection of high affineofpeptide ligands forcells detection 18 10 Separation of Escherichia coli of ClostridiumTyrobutyricumspores
Magnetic ion-exchange for medical, biochemical and molecular biological applications 20 Proteome of Human Serum Drug Based on Magnetic ClinicalProfiling Applications of Magnetic Targeting Purification of endotoxins from protein solution11 19 Bead Separation12
Enrichment of glyoproteins13
Introduction: Magnetic beads are becoming more and more popular in diverse applications for separation, concentration and purification of biomolecules. Applications like nucleic acid isolations, nucleic acid fragment size selection, PCR and ligation clean ups, protein purifications, specific cells or cell compartments depend upon the magnetic bead surface coatings and the interactive properties of the ligand molecules. Commonly used ligands include streptavidin, protein A, protein G, biotinylated antibodies, silica, COOH, SH, CHO and C18-C14.
Binding assays with streptavidin14 Immobilization of l-lactate dehydrogenase on magnetic nanoclusters for synthesis of pharmaceutical compounds15 Functional Selection of Vaccine Candidate Peptides from Staphylococcus aureus Whole-Genome Expression Libraries In Vitro16 Magnetic isolation of circulating tumor cells17 Separation of Escherichia coli18 Magnetic ion-exchange for medical, biochemical and molecular biological applications19 Clinical Applications of Magnetic Drug Targeting20 Figure 1 (A). Design of automated bead mixer (B). Bead suspension on the automated prototype bead mixer on the deck of VERSA Mini NAP/PCR Setup Workstation
A
(A). Design of automated bead mixer (B). Bead suspension on the automated prototype bead mixer on the deck of VERSA Mini NAP/PCR Setup Workstation
Figure 2
B
Standard curve of the magnetic beads suspension prepared by manual suspension (n=1)
0.25 0.2
OD600
Figure 1
0.15 OD600 0.1 0.05 0 128
64
32
16
8
4
2
1
Dilution (Fold)
Figure 2: Standard curve of the magnetic beads suspension prepared by manual suspension (n=1)
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July 2011
feature
g
In these applications, a uniform suspension and distribution of the beads is important. Therefore an efficient and non-magnetic method, such as a bead mixer, is required to keep the beads uniformly suspended in the source tube or reservoir. An efficient bead mixer will only ensure uniform distribution to the target wells of the multi-well plate or tubes if the beads are constantly kept in uniform suspension in the source tube or reservoir. At the same time, it is also important that the source tube or reservoir must not have “dead zones” or “fall out zones” where the beads fall out of solution. The “fall out zone” problem is very prominent in bead mixers that use propeller blade stirrers or ultrasonics. Since magnetic beads are very precious reagents, the bead mixer should minimize wastage of the bead solutions that occurs during pipette mixing of the beads. The latter results in beads sticking inside the pipette tips are due to high aspiration volumes required for proper mixing. Bead mixers using sealed drive shafts also pose problems due to wear and tear of the bearings and seals. On the other hand beads tend to collect in pockets between the bearings and seal. However, stirring of magnetic beads is also very difficult with bead mixers that use magnetic fields since the beads get attracted to such magnetic sources. This results in beads being pulled out of solution. In the present study, a prototype automated magnetic bead mixer manufactured by Aurora Biomed Inc was validated.
manually drawn from top, middle and bottom part of the tube containing the bead suspension. c. The distribution of the beads (20 µL) was carried from the bead suspension in source tubes to the wells
of a 96-well multi-well plate using VERSA Mini NAP/PCR Setup Workstation (Aurora Biomed Inc, Vancouver, BC, Canada). This sampling was carried out in replicates (n=10) to determine the unifor-
mity of distribution of the beads. The beads from each well of the target plate were re-suspended in 1 mL of distilled water to read OD600 on the spectrophotometer. d. A parallel sampling dis-
Materials & Methods: a. The magnetic beads were suspended in a source tube (outer diameter 1.4 cm x height 5.5 cm) to make a final volume of 5 mL in lysis solution used for nucleic acid isolation. If not otherwise stated, aliquots of 20 µL magnetic beads suspension were suspended in 1 ml of distilled water to read OD600 on a Spectrumlab 22PC spectrophotometer (Spectrumlab, Shanghai, PRC). b. The tube containing the beads was inserted into the bead mixer. Samples were
Reply card #4454
tribution was performed using a manual pipettor to sample the bead suspension following manual vortexing of the source tube to ensure optimum suspension.
0.18
10
OD 600 Mean (n=3)
0.16
feAture
0.14 0.12
Automation
0.1
Manual
July 2011 Laboratory Focus www.bioscienceworld.ca
0.08 0.06 0.04 0.02 0
Top
0.2
0.2
0.18
0.18
0.16
0.16
0.14
0.14
0.12
Automation
0.1
Manual
0.08
Mid
Bottom
Automated distribution of the magnetic bead suspension (automated mixing) to the target well figure Figure 3: Bar4diagram the quality of suspension of magnetic beads ofshowing the multi-well plate for nucleic extraction or in the source tube (Manually mixing versus automated mixing) (n=3) purification (n=1)
OD600 (n=1)
OD 600 Mean (n=3)
figure 3
bar diagram showing the quality of suspension of magnetic beads in the source tube (Manually mixing versus automated mixing) (n=3)
0.12 0.1
OD600
0.08 0.06
0.06
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0.04
0.02
0.02
0
0 Top
Mid
1
Bottom
2
3
4
5
6
7
8
9
10
Well Num ber
Figure 3: Bar diagram showing the quality of suspension of magnetic beads in the source tube (Manually mixing versus automated mixing) (n=3) Figure 4: Automated distribution of the magnetic bead suspension (automated mixing) to the target well of the multi-well plate for nucleic extraction or purification (n=1)
Results0.2and Discussion: The bead 0.18 mixer (Figure 1) was found to meet the re0.16 quirements of an ideal bead 0.14 mixer. A standard curve was 0.12 using the data generated from a range 0.1 of bead suspenOD600 sions manually mixed (Fig0.08 ure 2). The bead mixer provided a0.06 uniform suspension 0.04 of beads in the source tube under the 0.02 bead mixing conditions used. This statement 0 is supported by data from the 1 2 3 4 5 6 7 8 9 10 chová L., Slováková OD600 values obtained from 2006; 72(8): 5150–5158. Well Num ber M., Le-Nell A., Chmelik the top, middle and bottom 4. Sureselect:Anna KiialJ., Foret F., Viovy J-L., part of the source tube (CV% ainen, Olof Karlberg, Bilková Z.. J. Sep. Sci. 1.0 to 3.6). The uniformity of Annika Ahlford, SnaeFigure 4: Automated distribution of the magnetic bead suspension (automated to –the 2008;mixing) 31: 507 515. suspension was equivalent var Sigurdsson, Kerstin target well of the multi-well plate for nucleic extraction or purification (n=1) 10. Lavilla M. Ruth De Luis, for both automated and manLindblad-Toh, and AnnOD600 (n=1)
bead mixers using sealed drive shafts also pose problems due to wear and tear of the bearings and seals. On the other hand beads tend to collect in pockets between the bearings and seal.
ual mixing (Figure 3). The distribution of the beads taken from the automated suspension in the source tube to the target wells of the multiwell plate to be used for nucleic acid extraction or some other genomic applications, was found to be smooth and uniform (Figure 4).
References: 1. Berensmeier S. Appl Microbiol Biotechnol. 2006, 73:495–504. 2. Perelle S, Cavellini L, Burger C, Blaise-Boisseau S, Hennechart-Collette C, Merle G, Fach P. J Virol Methods. 2009 Apr;157(1):80-3. 3. Stratmann J., Karen Dohmann, Julia Heinzmann, and GeraldF. Gerlach Applied Environmental Microbiology
5.
6.
7.
8.
9.
Christine Syvänen. PLoS One. 2011; 6(2): e16486. Sébastien Rodrigue,# Arne C. Materna,# Sonia C. Timberlake, Matthew C. Blackburn, Rex R. Malmstrom, Eric J. Alm,* and Sallie W. Chisholm* PLoS One. 2010; 5(7): e11840. Liu P, Li X, Greenspoon SA, Scherer JR, Mathies RA. Lab Chip. 2011 Mar 21;11(6):1041-8. Frenzel A., Bergemann C., Köhl G., Reinard T. J. Chromatography B 2003; 793: 325-329. Pino P., Almudena Munoz-Javier, Dialechti Vlaskou, Pilar Rivera Gil, Christian Plank and Wolfgang J. Parak. Nano Lett., 2010. Korecká L., Jankovicová B., Krenková J., Herny-
María Dolores Pérez, Miguel Calvo, Lourdes Sánchez. Journal of Microbiological Methods 2009; 79: 214–219. 11. Asgar E., Büssow K., Oster J., Sieper J., Thiel A., Duchmann R. and Adam T. Microbial Cell Factories 2007; 6:18. 12. Baumann S., Uta Ceglarek, Georg Martin Fiedler, Jan Lembcke, Alexander Leichtle, and Joachim Thiery Clinical Chemistry 2005; 51(6): 973-980. 13. Tang Jia, Yang Liu, Peng Yin, Guoping Yao, Guoquan Yan, Chunhui Deng, Xiangmin Zhang Proteomics 2010; 10(10): 2000-2014. 14. Heim E., Frank Ludwig, Meinhard Schilling J. of Magnetism and Magnet-
ic Materials 2009; 321: 1628-1631. 15. Yusdy, Sohan R. Patel, Miranda G.S. Yapa, Daniel I.C. Wanga. Biochemical Engineering Journal Volume 48, Issue 1, 15 December 2009, Pages information. 16. Thomas Weichhart, Markus Horky, Johannes Söllner, Susanne Gangl, Tamàs Henics, Eszter Nagy, Andreas Meinke, Alexander von Gabain, Claire M. Fraser, Steve R. Gill, Martin Hafner, and Uwe von Ahsen. Infect Immun. 2003 August; 71(8): 4633–4641. 17. Sieben S., Bergemann C., Lübbe A.S., Brockmann B., Rescheleit D. J. of Magnetism and Magnet-
ic Materials 2001; 225: 175-179. 18. Deponte S., Steingroewer J., Löser C., Boschke E. and Bley T.. Anal. Bioanal. Chem. 2004; 379: 419-426. 19. Bergemann C., MüllerSchulte D., Oster J., Brassard L., Lübbe A.S.. J. of Magnetism and Magnetic Materials 1999; 194: 45-52 20. Lübbe A.S., M.D., Ph.D., Alexiou C. and Bergemann C.. J. of Surgical Res. 2001; 95: 200-206.
Acknowledgement: The authors would like to thank Saranna Brugger for editing the manuscript for final publication.
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feature
B y V i v e k J os h i , P h D , S e n i o r S c i e n t i s t , B i osc i e nc e B u s i n e ss Un i t * an d Es t e ll e R i c h e , S e n i o r S c i e n t i s t , L ab Wa t e r B u s i n e ss F i e l d , E M D M i ll i po r e
Optimizing Ultra High Performance Liquid Chromatography
Best Practices for Sample and Mobile Phase Preparation throughput of chromatography systems. This article will describe best practices that help ensure proper and efficient sample and mobile phase preparation for UHPLC analyses.
Table 1
Options for sample filtration
Sample Filtration Devices Sample Preparation Formats
Sample Preparation In UHPLC, sample clarification and fine particle removal are essential steps in preventing column clogging and system downtime. Some other sample preparation techniques (solid-phase extraction and liquid-liquid extraction) also reduce sample complexity, which leads to higher signal-to-noise ratios and cleaner baselines. Samples must be free of interfering matrix components that can bind the stationary phase in the UHPLC column. Depending on the type and number of samples, a range
SPE Sample Attributes
Ultra high performance chromatography (UHPLC) offers well-recognized benefits. Improvements in speed and resolution of 30 to 50 per cent over standard HPLC separations are typical. Chromatography runs that had taken 5 to 20 minutes now require only a fraction of that time, and the increased resolution allows for better separation of components. Along with the greater speed and resolution of UHPLC come increased demands on sample and mobile phase preparation. Smaller packing material and tubing diameters make UHPLC systems highly susceptible to clogging by impurities. Selection of the optimal sample and mobile phase filtration strategy for UHPLC is critical to avoid downtime and ensure quality results. With extremely low detection limits, attention must also be focused on the quality of the reagents. Ultrapure water is the largest component of the mobile phase, and it plays an essential role for running blanks, diluting samples, mixing buffers, and preparing standards. The water used for UHPLC analyses must be of the highest quality to ensure accurate, reproducible results. The speed of UHPLC also puts great demand on the throughput of processes used for sample preparation. Commonly used sample prep filtration devices such as syringe filters are tedious, slow, and unable to keep pace with the higher
High-throughput
•
Filter Plates
Syringe Filters
Centrifugals
1 – 100 mL
< 2 mL
•
• •
•
Volume
10 µL – 10 mL
< 2 mL
Reducing Simple Complexity
•
• •
Clarification
of preparation methods can be chosen for UHPLC sample preparation, including sample filtration using 0.2 µm filters; protein precipitation followed by centrifugation or
Parallel filtration of chromatography samples using the Samplicity Filtration System (EMD Millipore).
Figure 1
Filtration of 0.3 µm polystyrene latex suspension using various 0.2 µm syringe filters shows characteristic differences between membrane retention.
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Feature Figure 3
Effect of filtration of different solvents on UHPLC system back-pressure. Following filtration of water and methanol (A) or ammonium acetate and acetonitrile (B), hydrophilic PTFE membranes still show the best backpressure performance in an UHPLC run followed by nylon and polypropylene disc filters.
filtration; solid phase extraction; ultrafiltration; or liquid-liquid extraction (Table 1). When using sample filtration techniques, it is important to understand that the retention characteristics of
Continuing
syringe filters can vary, and this variance can impact the quality of sample preparation. Figure 1 compares the retention characteristics of syringe filters from two different suppliers. The data shows that when compar-
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Mobile Phase Preparation
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AD_Insertion_LF_switch_3.6235x4.735.indd 3
ing nylon and PES membranes, both products show similar particle retention (> 95 per cent ). In the case of PTFE and PVDF, membrane filters from Supplier A retain greater than 95 per cent particles, but the membrane filters from Supplier B only retain about 80 per cent of particles. The filters from Supplier B, which allow 20 per cent of the particles to pass through, may cause a column to clog. As the throughput of chromatography systems continues to improve, sample preparation techniques struggle to keep pace, becoming more of a bottleneck. Syringe filtration is a very simple sample preparation technique, but serial processing of samples can easily become the slowest and most tedious step in the entire analytical workflow. At the other end of the sample preparation spectrum are robotic systems, which can be expensive and offer too much capacity for labs that handle a few dozen samples per day. Recent innovations in sample processing such as the Samplicity Filtration System (Figure 2, EMD Millipore) allow multiple samples to be processed in parallel, offering a throughput capacity that is well-aligned with the needs of most labs. The system allows up to eight samples to be simultaneously vacuum filtered directly into LC vials in seconds.
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Unlike HPLC where mobile phase filtration requirements aren’t as stringent, typical UHPLC columns have small interstitial spaces, as well as 0.2 µm frits at the end of the columns to retain particles, which can get clogged. Filtering mobile phase com-
6/27/2011 9:52:08 AM
ponents with the optimal membrane filter helps protect UHPLC systems from particulate impurities that may clog and shut down the system. Most UHPLC suppliers recommend the use of filters with a 0.2 µm pore size for preparing the mobile phase. In a study conducted by EMD Millipore, three common mobile phase solvents (water, acetonitrile, and methanol) were used for mobile phase preparation and the effect of filtration on back-pressure of the UHPLC system was evaluated (Figure 3). Of the various membrane filters evaluated, hydrophilic polytetrafluoroethylene (PTFE) provided the best filtration performance as indicated by the lowest back-pressure increase in a UHPLC system. Hydrophilic polypropylene (PP) filters were unable to retain particulate impurities present in the solvents as indicated by highest backpressure gain of all the filters studied. Nylon and hydrophilic polyvinylidine fluoride (PVDF) filters show an intermediate performance in terms of particle retention and subsequent pressure increase. In addition to offering the lowest back pressure increase, hydrophilic PTFE filters also show the best chemical compatibility for various solvents and mobile phase modifiers commonly used in UHPLC (Table 2). It is also critical to use high quality buffer salts when preparing the mobile phase for UHPLC. Yet even with use of high quality salts, the presence of insoluble impurities can clog the UHPLC column. It is therefore important to filter mobile phase components prior to their use.
Water Quality Few factors affect high-performance liquid chromatography analyses more than contaminants in the water used for the mobile phase. While poor water quality is one of the easiest problems to fix, it is one of the least-understood factors in an analytical laboratory. Reports indicate that 70 to 80 per cent of chromatography performance issues are ultimately attributed to water quality in eluents, samples, and standards. Water contaminants that affect HPLC also impact UHPLC, but to a more significant extent in some cases. With detection limits down to partper-trillion levels, the water used for running blanks, diluting samples, mixing buffers, and preparing standards must be of the highest quality. Water used for UHPLC should be free of particulates, organic contaminants, bacterial contamination and ionic impurities. Particulates The presence of particulates in water can have a significant impact on UHPLC, due to its lower interstitial void
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feAture table 2
Membrane Chemical Compatibilities
Water
ACN
IPA
DMSO
DMF
0.2% FA
0.1% TEA
10 mM Phosphate
50 mM NH4OH
0.1% TFA
50 mM NH4OAc
Modifiers
MeOH
Diluents
Solvents and Buffers Membrane Materials
Mobile Phase
PTFE
E
E
E
E
E
E
E
E
E
E
E
E
PVDF
E
E
P
E
P
P
E
E
E
E
E
E
Nylon
G
E
E
G
E
E
P
E
E
N
P
E
PES
G
E
P
G
P
N
N
E
E
P
E
N
E = Excellent
G = Good
P = Poor
N = No Data
Membrane selection for mobile phase preparation. Hydrophilic PTFe disc filters show the best chemical compatibility of various membranes evaluated followed by PVDF, nylon and PeS.
volumes and decreased column diameters. Although UHPLC columns still operate at flow rates typical of conventional HPLC, the columns are more susceptible to premature plugging by particulates compared to HPLC columns. Organics Organic contamination of ultrapure water may affect chromatographic separations in a number of ways. Organic molecules may accumulate at the head of the column and subsequently elute as contaminant or ghost peaks. If the level of organic contamination is very high, the contaminants may act like a new stationary phase, causing shifts in retention time and peak tailing. Accumulation of organic material in the column may also lead to back pressure increases. This would ultimately shorten the column lifetime. In addition, the presence of organic contaminants in the water used to prepare the eluents may lead to a loss in resolution and reduced sensitivity of the analytical method. It is therefore critical to accurately monitor the level of organics in water used for UHPLC via an on-line TOC monitor. Bacteria Bacteria may plug column and frits and release organic by-
products which can, in turn, contribute to the effects seen with organic contamination. Ionic Impurities Modification of the ionic strength of the eluent may affect some separations. If the ionic contaminant is UVabsorbing (e.g., nitrates, nitrites), it will elute as a peak and make data analysis difficult. If a mass spectrometer (MS) is used as a detector, formation of adducts other than the protonated one (e.g., Na+, K+ adducts) can also impact data analysis.
Water purification systems combine several purification technologies to ensure that these important water contaminants are efficiently removed, and the highest water purity level is reached. A good pretreatment system combines reverse osmosis and electrodeionization technologies (such as in the Elix® system). Water is further purified with a polishing unit (e.g. a Milli-Q® system) combining UV photooxidation, ion exchange resins, activated carbon, as well as microfiltration at the point-of-use.
Water purification systems combine several purification technologies to ensure that these important water contaminants are efficiently removed, and the highest water purity level is reached.
The storage of ultrapure water in plastic containers can also impact UHPLC as the containers may introduce leachables into the water. Glass containers are preferred over plastic, as they do not leach as many organics, however they can introduce ions into the water. Storing ultrapure water can also encourage the proliferation of bacteria. Therefore, freshly purified ultrapure water should be preferred.
Conclusion The benefits of UHPLC are clear: higher speed, improved resolution, and increased throughput. In order to fully achieve these benefits, however, processes used for sample and mobile phase preparation must meet
higher standards that those used for HPLC applications. The use of the most appropriate filtration materials and sample prep technologies, as well as the highest quality water, all contribute to maximizing the effectiveness of UHPLC.
References: 1.
Mabic S., Regnault C., Krol J. The misunderstood laboratory solvent: reagent water for HPLC. LCGC North America 23(1):74-82 (2005). 2. Tarun M., Monferran C., Devaux C., Mabic S. Improving chromatographic performance by using freshly delivered ultrapure water in the mobile phase. LCGC „The Peak“, June, 7-14 (2009)
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New Products Spectrofluorometers The JASCO FP8000 Series spectrofluorometers incorporate high sensitivity (> 5000, RMS), fast spectral scanning capability (60,000 nm/ min) and excellent analysis-oriented functionality with integrated solutions for advanced materials research and biochemical analysis applications. A variety of accessories are available for integration with a range of sophisticated control and analysis applications available in the user-friendly Spectra Manager™ II software to offer a flexible platform for any luminescence application.
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Washer BioTek Instruments, Inc. new ELx405 Select Deep Well Washer is a robot compatible, full plate washer for automated aspirate and dispense steps in 96- and 384-well plates up to 50 mm high, and can also accommodate standard 15 mm microplates. The ELx405 also features BioTek’s patented Dual-Action™ manifold for independent and precise aspirate/dispense control, and variable flow rates and angled dispensing for optimized cell washing. Washing protocols may be created and run through the familiar keypad interface or through BioTek’s LHC™ Software. Additionally optional Ultrasonic Advantage™ integrated ultrasonic cleaners provide manifold self-maintenance, and optional automatic 1-4 wash buffer switching further enhances efficiency. Typical applications that can now be automated include: plasmid DNA purification, serum/plasma sample preparation, hematopoietic progenitor cell enumeration and cell proliferation assays.
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Tissue Slicer Warner Instruments introduces the OTS 5000 Automatic Oscillating Tissue Slicer for sectioning fixed or fresh tissue. Engineered for maximum stability, the OTS 5000 slicer head minimizes ZAxis vibration, allowing for even the most delicate tissue to be cut successfully every time. The OTS-5000 features a user programmable cutting window to minimize time between harvesting consecutive slices, allowing the section-cutting range of the blade to be customized to the specimen sample. Slice one section at a time, altering tissue thickness between manually-triggered passes. Or, automatically perform multiple uniform slices. The OTS-5000 is also available with a refrigerated controller and tray to maintain tissue temperature between 0-10°C.
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Incubator BINDER CB series incubators provide excellent growth conditions for mammalian cell and tissue cultures, offering precise, reproducible control of oxygen concentrations as low as 0.2 per cent and quick recovery times, important factors in conducting research under hypoxic conditions. Models CB 53, 150 and 210 (model numbers refer to chamber volume in liters) allow sample access via optional small internal doors (four on the 53 and 150, six on the 210) that, in addition to maintaining steady CO2 and O2 levels and accelerating recovery, minimize the risk of contamination. The incubators feature as standard a 180°C hot-air cycle that provides true sterilization. Interior walls stay dry through controlled humidification with a defined condensation point, and chamber preheating with the VENTAIR™ air jacket provides homogeneous temperature distribution. A seamless, deep-drawn interior features an integrated shelf-support system for easy cleaning.
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Transducer OMEGA introduces a new metric high accuracy all stainless steel pressure transducer. The PXM309 Metric series has an exceptional static accuracy of 0.25% per cent and a 1 per cent total error band (TEB) over a compensated range of -20 to 85°C (on ranges of one to 700 bar). Ranges from 70 mbar to 3.5 bar are also available with a slightly larger TEB. All units are supplied with a five-point NIST traceable calibration certificate. The all Stainless steel construction and solid state reliability make these transducers suitable for most industrial applications as well as engine test stands. The PXM309 Series transducers are sealed to an IP65 environmental rating and are available with three electrical terminations, a two meter cable with flying leads, a mini-DIN connector or a 4-pin M12 connector. Two outputs are available, either 0-10 Vdc or 4-20 mA. The pressure connection is a standard G ¼ male metric fitting.
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HPLC Column Range Thermo Fisher Scientific Inc. announces the release of the Thermo Scientific Accucore HPLC column range. Founded on core enhanced technology and utilizing phase bonding and packing, these columns provide a unique chromatography solution to enhance laboratory workflow and efficiency. Available in a wide range of stationary phase selectivities and compatible with almost any instrument, these columns give increased sensitivity and peak resolution. Containing solid core particles, which are engineered to a diameter of 2.6 µm and an average particle size distribution (D90/10) of 1.12, the Thermo Scientific Accucore HPLC column range allows high-speed, high-resolution separation, with backpressures significantly lower than those associated with UHPLC. The advanced bonding technology used in Accucore HPLC columns results in excellent peak shapes through high bonding coverage and minimized secondary interactions. The automated packing processes deployed in column manufacture produce uniform tightly packed columns, which are robust and reproducible.
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ElectroPrep System The new ElectroPrep system from Harvard Apparatus is an extremely versatile patented sample prep technology based on electro-dialysis. The ElectroPrep can be used for electrodialysis, electro-elution, electrofiltration, electro-fractionation or electro-concentration for running a sample simultaneously into multiple molecular weight cut off (MWCO) fractions in as little as three minutes. This system is ideal for the rapid purification of proteins, nucleic acids, carbohydrates and other biomolecules. With a run-time of three to 10 minutes, ElectroPrep provides speed and convenience, even at the very low currents (five to10mA) used with this system. The system’s reusable sample chambers are made of PTFE for high sample recovery and have been redesigned to provide larger membrane surface areas for even faster dialysis rates.
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Thermometer Control Company’s new Traceable® WideRange Big-Digit Refrigerator Thermometer has 7/8-inch high display that is easy to view. Switchable range is from –3.8 to 199.9°F and –19.9 to 110.0°C with a resolution of 0.1°. Accuracy is ±1ºC in the range 0 to 25°C and ±1.5°C outside range. Ultra™ model accuracy is ±0.5°C at tested points. The thermometer may be placed anywhere using four methods: Always-hold™ secure suction cup, place-anywhere Velcro®, stand, and hanger. Display updates every 30 seconds (this allows capturing a reading while allowing a momentary open refrigerator door).
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Separation-ESI Module The CESI 8000 High-Performance Separation-ESI Module with OptiMS Technology from Beckman Coulter, Inc. consists of the first commercial CESI sprayer combined with new capillary electrophoresis instrumentation specifically designed for mass spectrometry (MS). This gives researchers a robust, high-sensitivity interface of capillary electrophoresis (CE) with MS, while providing a broad range of analyte coverage complementary to LC-MS and GC-MS. The technology integrates low flow CE separation (as low as 10 nL/ min.) with electrospray ionization (ESI) in a single process that takes place within a capillary. The new process is referred to as CESI, reflecting the combination. CESI-MS is capable of generating separations of peptides with a high peak capacity (>320), allowing the analysis of very complex samples at the proteome level. The ultra-low flow rates used by CE effectively reduce ion suppression and make it possible to detect post-translational modifications (PTMs) like phosphorylation and sialylation, which typically hamper efficient ionization. High-separation efficiency and sensitivity permit the detection of peptides present in low abundance at concentrations in the picoMolar range. The selectivity of CE in front-end separation further enables the characterization of intact proteins, their isoforms, cleaved fragments and complexes. Easy switching between LC-ESI-MS and CESI-MS allows the addition of CESI to existing workflows.
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New Products Titrator JM Science announces its new AQUACOUNTER® Karl Fischer Volumetric Titrator (AQV-2200S). This new titrator is ideal for users who: 1) have many kinds of samples to be analyzed; 2) exchange KF reagents frequently or after each measurement. Small volume titration cell requires only 20mL of titration solvent for accurate measurements. Less reagent volume reduces waste and is easy to replace with fresh solvent for the next measurement. Combining one additional KF station with either a volumetric or coulometric module enables two different measurements to be performed at the same time. With both a volumetric and coulometric channel, one can measure moisture over the entire range from 1ppm to 100 per cent or double your sample throughput with 2 identical KF stations. A two-channel option means easy “plug-andplay”. End-users can add various peripherals, such as a second channel, and the system recognizes the new channels and begins working with them immediately. Current status, data and function keys are displayed on a colorful and large screen. Color touch screen is 7.5-inches wide. Result data with curves can be viewed on your PC internet browsing program without optional software.
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Pipette The new Thermo Scientific F1-ClipTip pipette and tip system is the latest addition to a new generation of Thermo Fisher Scientific ergonomic products. The F1-ClipTip™ system provides extremely light-touch dispensing and tip ejection for superior ergonomic performance. The manual liquid handling system includes all of the proven features of the Thermo Scientific F1 pipettes, but adds an innovative tip interface that takes pipetting to new levels of liquid dispensing consistency. ClipTip pipette tips have a hydrophobic inner surface to prevent liquid retention, while optional filter tips use the aerosol resistant tip (ART) self-sealing barrier to reduce cross-contamination. With the new F1-ClipTip pipette and tip system, users can clip it and forget it. It’s that simple. As a complete system for secure pipetting, the pipettes and tips are available in a variety of formats or as a Good Laboratory Pipetting (GLP) Kit.
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Pump Valco Instruments new generation of liquid handling pumps are designed for precision handling of liquids and/ or gases, producing a bidirectional pulseless flow with a range of over six orders of magnitude (5 nL/min to 5 mL/min or 1 µL/min to 25 mL/ min, depending on model). Manufacturer says the pumps offer better performance than syringe pumps and eliminates the need for refill cycles and syringe changes. The positive displacement pumps are self-priming and tolerant of any gas which may find its way into the fluid lines. There is no separate fill cycle, and the capacity is unlimited.
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rotary control knobs or a keypad. A quick pre-cooling function allows the centrifuge’s temperature to go from 21ºC to 4ºC in only eight minutes. The centrifuge has been developed to meet latest emission and ecological requirements.
Eppendorf’s refrigerated 24-place microcentrifuge comes equipped with many innovative and usable design features, and has won a red dot: product design 2011 award for design excellence in the life science and medicine category. With its very compact size, low access height and near-silent operation - even without the rotor lid the centrifuge fits easily into molecular and biological research laboratories. Four different rotors ensure versatility, while users can choose between
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Mobile Bench Labconco’s multi-use mobile bench provides bench space and transports equipment from lab to lab. It is made of durable and chemical-resistant
epoxy-coated 18-gauge welded steel frames of 5.1 cm square tubing and is painted with tough, corrosion-resistant, dry powder epoxy in glacier white. The bench also features 12.7 cm diameter, non-marking polyurethane casters that absorb vibration over rough, uneven floors for stability and quiet operation. The work surface is fiber board with high pressure melamine laminate that is 2.8 cm thick. The bench is strong and supports loads up to 245 kg.
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JULY July 26-30 ASCLS Annual Meeting Venue: Atlanta, GA Tel: 301-657-2768 Fax: 301-657-2909 Email: ascls@ascls.org Web: www.ascls.org
July 27-29 International Conference on Nanotechnology Venue: Ottawa, ON Email: Info@ International-ASET.com Web: http://icnfa2011. international-aset.com
July 2011
September 27-28 HSE Canada 2011 Venue: Toronto, ON Tel: 647-893-8984 Email: nate@ hsecanada.ca Web: www.hsecanada.ca
September 29-October 1 BCSLS Congress 2011 Venue: Castlegar, BC
Calendar Tel: 604-714-1760 Fax: 604-738-4080 Email: bcsls@telus.net Web: www.bcsls.net
OCTOBER October 5-6
NOVEMBER November 16-19
BioContact Quebec Venue: Québec, QC Web: www.biocontact.ca
Germany Web: www.medica.de
November 28-30
Medica-World Forum for Medicine Venue: Dusseldorf,
Canada Renewable Fuels Summit Venue: Calgary, AB Web: www.greenfuels.org
AUGUST August 7-11 Microscopy and Microanalysis 2011 Venue: Nashville, TN Tel: 703-234-4115 Fax: 703-435-4390 Email: Association Management@microscopy.org Web: www.microscopy.org
August 28-31 ICASS and CCACC 2011 Symposia Venue: Toronto, ON Fax: 905-589-0059 Email: janette@ etpsymposium.com Website: www.csass.org
SEPTEMBER September 3 International Pain Summit Venue: Montreal, QC Tel: 206-283-0311 Fax: 206-283-9403 Email: iaspdesk@iasp-pain.org Web: www.iasp-pain.org
September 7-10 World Molecular Imaging Congress Venue: San Diego, CA Tel: 310-215-9730 Fax: 310-215-9731 Email: ami@ami-imaging.org Web: www.ami-imaging.org
September 13-15 Stem Cells USA & Regenerative Medicine Congress Venue: Boston, MA Tel: 212-379-6322 Fax: 212-379-6319 Email: enquiry.us@ terrapinn.com Web: www.terrapinn.com
September 16-23 National Biotech Week Web: www.imagenenation.ca
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July 2011 Laboratory Focus
Toronto welcomes back the largest international meeting of of stem Cell researchers The International Society for Stem Cell Research (ISSCR) 9th Annual meeting, the largest international interdisciplinary forum dedicated to stem cell science, made its return to the Metro Toronto Convention Centre (MTCC). Running June 14-19, the event attracted some 3,500 researchers from academia and industry to discuss emerging science in the field. Attendees were treated to outstanding speakers, a huge floor show and lecture schedule that was packed with the best and brightest the stem cell field had to offer. The latest discoveries in this rapidly moving field were on full display as 111 speakers from academia and industry gave presentations while the floor show featured more than 1,400 poster abstracts and 132 exhibitors. Three key awards – the ISSCR McEwen Centre Award for Innovation, the ISSCR Public Service Award and the Outstanding Young Investigator Award – were also handed out honouring some of the best researchers in the field. The recipients of the inaugural ISSCR McEwen Centre Award for Innovation were Kazutoshi Takahashi and Shinya Yamanaka. The award recognized their paradigm-shifting work demonstrating the reprogramming of adult/tissue-specific cells using transcription factors that has resulted in a rapid development of novel tools and strategies for use in the pursuit of better understanding and treating disease. The inaugural ISSCR Public Service Award was presented to Robert Klein, chairman of the Governing Board of the California Institute for Regenerative Medicine (CIRM), for his outstanding contribution of public service to the field of stem cell research and regenerative medicine, while UCSF’s Robert Blelloch, MD, PhD, received the 2011 Outstanding Young Investigator Award from the International Society for Stem Cell Research, for his pioneering research on the role of molecular tools known as microRNAs in embryonic stem cells and cancer. On the Canadian front, a community of local scientists used the ISSCR to launch an online portal to open the world of stem cells to the public. This virtual community, called Stem Cell City, is designed to teach people about stem cells and to inform them of the latest research aimed at defeating the most devastating diseases afflicting humans. In other exciting Canadian news, a new partnership linking sixty-five stem cell research laboratories from across Ontario known as the Ontario Stem Cell Initiative (OSCI), officially launched. Related to this, OSCI members also announced the launch of The Centre for Commercialization of Regenerative Medicine (CCRM), a new centre to drive commercialization of regenerative medicine and attract millions of dollars to Ontario. It was fitting that on the 50th anniversary of Canadian researchers Drs James Till and the late Dr. Ernest McCulloch’s first published a paper proving the existence of stem cells, that the event was hosted in the city where the milestone occurred. While there are no immediate plans to bring the event back to Toronto, organizers agreed that it should make another return sometime soon. Already a date and time has been set for both next year and 2013. The 10th annual ISSCR meeting will take place June 13-16 in Yokohama, Japan. It will return to North America in 2013, June 12-15 in Boston.
CAreer SPOtLight Bio-economy Career Profile Compiled by BioTalent Canada Position: Bio-Strategy Consultant Name: Andrew Hessel Company: Alberta Ingenuity Fund (AIF)
What I do:
I promote synthetic technology in Alberta and beyond. AIF is a $1 billion fund that supports and encourages development in the areas of science, technology, and engineering. I focus my efforts in digital biology, founded on machines that print and assemble DNA code. The new technology allows software-based DNA manipulation to write new genetic programs easily for a broad range of applications. I work at three different levels for AIF in promoting the new technologies and by creating greater awareness of their applications. First, I develop outreach programs designed for students from high school to postgrad. Second, I work with universities to create courses and degree programs in digital genetic engineering. Finally, I work to raise awareness about the technology to science- funding and industry groups about the changing dynamics and economics of genetic engineering.
What education and skills do candidates need for this position?
My educational background includes a Bachelor of Cellular, Molecular and Microbial Biology, a Masters in Bacterial Genomics, and years of networking with leading scientists and engineers. You definitely require a strong scientific foundation combined with a broader understanding. Strong computer skills and experience using software tools are also required, as digital information and communication are key to modern life science. Social networking is becoming more important, as people working in digital biology need to collaborate and share data with others from around the world. An understanding of business and the use of intellectual property are crucial assets, since commercialization of technology is necessary for development and sustainable research funding. It’s essential to have a clear vision and the ability to communicate it effectively. You need to challenge convention and be willing to be disruptive at times. Many industries and their corporations have an institutional inertia that sometimes does not allow them to look beyond their organizations or rapidly commit to change. As a bio-strategy consultant, you need to always be looking for ways to improve what is already out there, and to re-evaluate and be willing to change conventional wisdom. A certain type of stubbornness is needed. Don’t be afraid to break out of your comfort zone and look in some very different directions, whether it’s an MBA, finance, or public speaking.
Y O B
T c S s p o w
What are the best parts of your job?
The best thing about my job is that I connect with brilliant and empowered people from around the world, who look toward the future for the betterment of all. I work with people from academia, business, politics, and finance, who are interested in making fundamental change to existing systems and improving them.
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