Laboratory Focus November/December 2016 by Promotive Communications

Pharmaceutical

Clinical

Chemical

w w w. l a b o r a t o r y f o c u s . c a Paramagnetic Bead Based Nucleic Acid Isolation Page 8

food

environment

November/December 2016 Volume 20, Number 4

Liquid Transfer in Cell Culture with Multipette® M4 Repeater® M4 Page 14

R&D News.......................... 1 Appointments..................... 6 Pharma Notes..................... 7 New Products................... 16 Calendar........................... 17 App Reviews...................... 18

New U of S immunotherapy technique holds promise for curing food allergies SASKATOON, SK – University of Saskatchewan (U of S) scientists have developed a new immunotherapy technique that nearly eliminates the allergic response to peanut and egg white proteins in food-allergic mice, reducing the anaphylactic response by up to 90 per cent with only one treatment. “This discovery reverses food allergies in mice, and we have many people with allergies volunteering their own cells for us to use in lab testing to move this research forward,” says professor John Gordon, lead scientist behind the discovery that appears in the current issue of the Journal of Allergy and Clinical Immunology. The findings open the door to test the treatment in “humanized mice” — mice with non-existent immune systems implanted with cells from a human immune system, for example, from a peanut-allergic person. With Health Canada approval, the first human trial could begin in about one year, Gordon says. “If we can reliably ‘cure’ food allergies, or related conditions such as asthma or autoimmune diseases such as multiple sclerosis with this new therapy, it would be lifechanging for affected individuals.” The discovery involves generating

Publications Mail Registration Number: 40052410

a type of naturally occurring immune cell that sends a signal to reverse the hyper-immune response present in allergic reactions. That signal triggers another “off switch” that turns off reactive cells further along the allergic pathway. “We predict the treatment could be on the market within the next five to 10 years,” says Gordon, who is also a research leader in the Allergy, Genes and Environment (AllerGen) Network. AllerGen—part of the federally funded Networks of Centres of Excellence program — aims to help Canadians address the challenges of living with asthma, allergies, anaphylaxis and related immune diseases. Gordon’s team will collaborate with other AllerGen investigators located at the U of S, McGill University, Queen’s University, McMaster University, and University of Alberta to pilot the new technique. There is already compelling evidence this technique could be effective in humans. In 2010, Gordon’s team demonstrated they could reverse an asthmatic response in human cells in a test tube. Using three applications of a similar therapy in a 2012 study, the researchers effectively eliminated asthma in afflicted mice, within only eight weeks.

Lead scientist John Gordon (foreground) and Wojciech Dawicki (background), first author of the article, in Gordon’s lab at the U of S. (Photo credit: University of Saskatchewan)

“Even if we only cure 25 per cent of subjects, we will dramatically improve the health of those individuals, and also reduce healthcare system expenses,” said Gordon, who worked with Wojciech Dawicki, a research associate and the primary author and lead researcher in this study. Master’s student Chunyan Li and lab technicians Xiaobei Zhang and Jennifer Town also worked on the project. Gordon said the new technique also shows promise for treating

autoimmune disorders such as multiple sclerosis. “It would take very little to adapt the therapy for autoimmune diseases,” he said. Funding for the research was provided by the Canadian Institutes of Health Research and the AllerGen Networks of Centres of Excellence. To see this story online visit http://www.laboratoryfocus.ca/ new-u-of-s-immunotherapy-technique-holds-promise-for-curingfood-allergies/

November/December 2016 Laboratory Focus www.laboratoryfocus.ca

news

Health organizations in Canada and China team up to accelerate precision medicine VANCOUVER, BC & SHENZHEN, CHINA – Five leading health and research organizations in British Columbia and the world’s largest genom-

ics organization, based in China, have formed a global partnership to bring together advanced technologies and individualized therapy for cancer

and other diseases. The partnership, between Shenzhen-based BGI, Providence Health Care, St. Paul’s Foundation, Vancouver Pros-

tate Centre (VPC), Genome British Columbia (Genome BC), and Deloitte Inc., foContinued on page 3

PUBLISHER/EDITOR-IN-CHIEF Terri Pavelic SENIOR WRITER Shawn Lawrence CONTRIBUTING WRITERS Alexander Stroganov Brian Rivera Diana Hübler Maxim Slyadnev Muriel Art Nathalie Chandelier Sikander Gill Vincent Dufey GRAPHIC DESIGNER Elena Pankova CONTROLLER John R. Jones MARKETING MANAGER Mary Malofy CIRCULATION DIRECTOR Mary Labao mary@promotivemedia.ca Tel: 905-841-7389

BDH® ANALYTICAL CHEMICALS CATALOG Order Your Copy Today! Visit vwr.com/BDHCatalog.

OFFICE: 23-4 Vata Court Aurora, ON L4G 4B6 Phone: 905-727-3875 Fax: 905-727-4428 E-mail: laboratory_focus@ promotivemedia.ca SUBSCRIPTION INQUIRIES mary@promotivemedia.ca Fax: 905-727-4428 Laboratory Focus is published 4 times per year by Promotive Communications Inc. Legal Depository: National Library of Canada ISSN 40052410 Subscription rate in Canada $35/year; USA $60/year; other countries $100/year. All rights reserved. No part of this publication may be reproduced without written consent. Publications Mail Registration Number: 40052410 Return undeliverable Canadian addresses to circulation dept: 23-4 Vata Court Aurora, ON L4G 4B6 E-mail: mary@promotivemedia.ca

NEW

All opinions expressed herein are those of the contributors and do not necessarily reflect the views of the publisher or any person or organization associated with the magazine. If you would like to order hard copy or electronic reprints of articles, contact

Includes HiPerSolv CHROMANORM® HPLC Solvents

www.laboratoryfocus.ca

twitter.com/@LabFocus linkedin.com/company/ laboratory-focus facebook.com/LaboratoryFocus

ocus

www.laboratoryfocus.ca Laboratory Focus

November/December 2016

news

Continued from page 2 cuses on applying next generation sequencing technologies and big data to accelerate personalized medicine and improve outcomes for patients. “This partnership links the best researchers in British Columbia and China with cutting edge genomic technologies to share expertise and accelerate innovation on a global scale,” said Dianne Doyle, president and CEO, Providence Health Care. “Our mutual goal is more effective treatments for patients using precision medicine.” The partners hope to leverage eachothers strengths to create health centres of excellence in both Shenzhen and Vancouver. For example, the partners envision creating a joint research centre at the new St. Paul’s Hospital campus that would involve scientists and clinicians from UBC’s

Faculty of Medicine and use BGI’s cost-effective, high-throughput sequencing capabilities. Deloitte will contribute its expertise managing the big data analytics environment to facilitate the research and enable health information to be securely shared with the various research partners. “BGI and BC have collaborated for many years,” said Ning Li, chief development officer of precision medicine at BGI. “This is an exciting step forward to build stronger collaboration to realize the goal of precision medicine, accelerate innovation and make an impact on improving human health.” A Memorandum of Understanding was signed by the parties at a ceremony at the Vancouver Board of Trade during the American Society of Human Genetics 2016 annual meeting.

To see this story online visit http://www.laboratoryfocus.ca/health-organizations-in-canada-and-chinateam-up-to-accelerate-precision-medicine/

Canadian neuroscience leaders tap IBM Watson to pinpoint new drugs for Parkinson’s disease

Discovery Advisor Scott Spangler. Photo: Jon Simon, Feature Photo Service for IBM

TORONTO, ON – The Ontario Brain Institute (OBI) and the Movement Disorders Clinic (MDC) at Toronto’s University Health Network (UHN) have embarked on Canada’s first ever Parkinson’s disease research project using the recently launched IBM Watson for Drug Discovery. Watson is the first commercially available cognitive computing capability representing a new era in computing. The system, delivered through the cloud, analyzes high volumes of data, understands complex questions posed in natural language, and proposes evidence-based answers. Watson continuously learns, gaining in value and knowledge over time, from previous interactions. MDC researchers, along with members of the Informatics and Analytics

team at OBI, will use Watson to accelerate the drug discovery process and determine which drugs could potentially be re-purposed in the fight against Parkinson’s disease. Drawing from its body of nearly 31 million sources of relevant literature, IBM’s cloud-based cognitive enterprise solution analyzes scientific knowledge and data using machine learning and natural language processing. According to the Parkinson’s Disease Foundation, an estimated seven to 10 million people globally are living with Parkinson’s disease, and currently it is the second most common neurodegenerative disorder in Canada, trailing only Alzheimer’s disease. Currently, bringing a drug to market takes nearly 10 years and approximate-

ly $2.6 billion. Beyond that, 88 per cent of new drugs fail in Phase 1 because of a lack of efficacy and safety. “Drug researchers are challenged by the sheer volume and pace of emerging data,” said Lauren O’Donnell, vice president, IBM Watson Health Life Sciences. “Watson for Drug Discovery empowers researchers with cognitive tools that will help to speed drug discovery, and increase the likelihood of bringing effective therapies to patients more rapidly.” “This partnership signals the beginning of a new era for neuroscience where researchers can work with data at an unprecedented level of sophistication and speed,” adds Tom Mikkelsen, president and scientific director of the Ontario Brain Institute. “We are excited by the impact this could have on people living with Parkinson’s disease.” Dr. Lorraine Kalia, a movement disorders neurologist and neuroscientist at the Krembil Neuroscience Centre at UHN, agrees that cognitive technology like IBM Watson has the potential to make discoveries that can directly impact the health of Canadians. “The platform gives us the ability to look at connections that researchers might not have found without dedicating weeks or months of time,” said Dr. Kalia. “This includes identifying compounds that we have not previously considered investigating for the treatment of Parkinson’s disease.” To see this story online visit http://www.laboratoryfocus.ca/ canadian-neuroscience-leaders-tapibm-watson-to-pinpoint-new-drugsfor-parkinsons-disease/

Canadian Court of Appeal upholds genetically modified salmon approval OTTAWA, ON – The Federal Court of Appeal in Canada has upheld the approval of genetically modified salmon produced by AquaBounty Technologies Inc, after dismissing an appeal brought by ecological organizations. Health Canada and the Canadian Food Inspection Agency had approved AquAdvantage Salmon for production, sale and consumption in Canada in May. The Federal Court of Appeal ruled that the Ministers had arrived at a decision that was “reasonable and made in the manner prescribed by the Canadian Environmental Protection Act, 1999, SC 1999, c 33.” The appeal was brought up by Ecojustice on behalf of the Ecology Action Centre and Living Oceans Society against Canada’s Minister of the Environment and Climate Change, Minister of Health and the company. “We welcome the decision made by the Court of Appeal. We thank the Ministries of Canada who were extraordinarily thorough and transparent in the review and approval of AquaBounty’s applications,” said Dr. Ronald L. Stotish, CEO of AquaBounty. To see this story online visit http://www.laboratoryfocus. ca/canadian-court-of-appealupholds-genetically-modifiedsalmon-approval/

November/December 2016 Laboratory Focus www.laboratoryfocus.ca

news

A metabolic switch to turn off obesity MONTRéAL, QC – You’ve tried all the diets. No matter: you’ve still regained the weight you lost, even when you eat well and exercise regularly. This may be due to a particular enzyme in the brain: the alpha/beta hydrolase domain-6 enzyme, better known as ABHD6. A study published in Cell Reports demonstrates that when this enzyme is blocked in certain neurons of the mouse hypothalamus, it becomes impossible for them to lose weight, even if they adhere to an ideal regimen, ideal for mice that is! The study, conducted by a research team at the University of Montréal Hospital Research Centre (CRCHUM) involved genetically engineered mice, deprived of the ABHD6 enzyme in a localized area of the brain, namely in a specific population of hypothalamic neurons. Alexandre Fisette, postdoctoral researcher at CRCHUM and first author of the study, explains that, “under normal conditions of housing and food, these mice are identical to normal mice. However, when challenged, they are unable to adapt. They no longer consume food after a fast, they cannot maintain their body temperature during exposure to cold, and they are more susceptible to be-

come obese when fed a high-fat diet. What’s more, once they are obese and we try to make them lose weight by feeding them a normal diet, they do not lose weight.” The researchers have discovered that this enzyme acts as a sort of switch for the body’s adjustment to extremes. “It is a mechanism we had not suspected. Strikingly, the absence of one single enzyme within a precise region of the brain completely disrupts the normal metabolism and prevents the mice from losing weight,” comments Thierry Alquier, CRCHUM researcher

and professor at the University of Montréal. Is there an identical process taking place in humans? Alquier thinks that clinical studies will be required to find out. However, according to Alquier, ABHD6 has a key role in the rebound effect that is often observed after a dietary regimen. “People who experience difficulty losing weight might have a deficiency of this enzyme,” he says. The pursuit of the ABHD6 enzyme appears to be promising. In 2014, the team of Marc Prentki, another CRCHUM researcher, discovered that this enzyme breaks down endocannabi-

noids. Blocking ABHD6 in peripheral organs and adipose tissues protects against obesity and against type 2 diabetes. “We know today that ABHD6 plays a completely different role in certain neurons of the hypothalamus. Blocking the enzyme in this location promotes obesity, whereas blocking it elsewhere in the body has beneficial effects,” emphasizes Stephanie Fulton, CRCHUM researcher and study co-author. Multiple signals and neuronal networks are involved in regulating the balance of energy so as to maintain a stable body weight. “We have shown the critical part played by the ABHD6 enzyme in preserving homeostasis in specific neurons of the hypothalamus. But we don’t know what happens when we block the enzyme in the entire brain. This is what we are currently investigating in an ongoing study,” says Fulton. The study was sponsored by the Canadian Institutes of Health Research (CIHR). To see this story online visit http://www.laboratoryfocus. ca/a-metabolic-switch-to-turn-offobesity/

New findings challenge current view of how pancreatic cancer develops

(L to R) Dr. Faiyaz Notta & Dr. Steven Gallinger, researchers in the multidisciplinary PanCuRx research initiative TORONTO, ON – Researchers in the multidisciplinary PanCuRx research initiative at the Ontario Institute for Cancer Research (OICR) and University Health Network’s Princess Margaret Cancer Centre, led by Dr. Faiyaz Notta and Dr. Steven Gallinger, have published new findings that challenge current

beliefs about how and why pancreatic cancer is so aggressive. The deadly disease often strikes “out of nowhere,” and the cancer is often inoperable by the time the patient experiences symptoms. Moreover, pancreatic cancer’s tendency to appear at an advanced stage has long been a medical mystery.

Using whole genome sequencing, the team reconstructed the history of pancreatic cancer development in 100 independent tumours. Unexpectedly, they found that many of the important alterations that are thought to cause this disease actually occur “all at once”, similar to the idea of the “big bang”. The results were published in the journal Nature. “When we started this project many years ago, we wanted to make better sense of how this disease progresses clinically,” said Dr. Notta, an OICR Fellow, principal investigator at the Princess Margaret and lead author of the study. “This disease can go from being a local cancer, restricted to the pancreas, to becoming fully metastatic very rapidly. The traditional view of the biology of the disease just didn’t jive with what happens clinically. And it’s hard to move forward in trying to find new treatments if you can’t link the biology of the tumour to the clinical reality of the disease. Our findings show a very different path for how this disease develops and puts the clinical problem of this disease into better perspective. We can make more sense about why this disease is

so aggressive and can advance so quickly.” “Pancreatic cancer is one of the most deadly types of cancer and still one of the least understood,” echoes Dr. Gallinger, head of Hepatobiliary/Pancreatic Surgical Oncology Program at UHN and Mount Sinai Hospital and leader of PanCuRx. “These findings provide us with a new understanding of how pancreatic cancer develops and a path forward to identify better strategies to diagnose and target this terrible disease.” Both researchers say that the new findings open up important new pathways of investigation that could lead to the ability to better diagnose pancreatic cancer, predict how it will develop and determine how and when it will metastasize. The findings could also be applicable to other aggressive tumour types. To see this story online visit http://www.laboratoryfocus.ca/ new-findings-challenge-currentview-of-how-pancreatic-cancerdevelops/

www.laboratoryfocus.ca

Laboratory Focus November/December 2016

SCPDC teams with Dana-Farber Cancer Institute on cancer research initiative

HAMILTON, ON – McMaster University’s Centre for Probe Development and Commercialization (CPDC) has entered into a new partnership with the Dana-Farber Cancer Institute that will see the two organizations working together to create novel molecular probes for non-invasive imaging in cancer research, drug development and patient care. The arising discoveries from this partnership could allow researchers and clinicians to use non-invasive molecular imaging to understand the

biology of cancers and accelerate the development of new therapeutics. Additionally, the two sides have formed an International Probe Development Consortium (IPDC) that will combine the technical expertise of the CPDC with Dana-Farber’s deep cancer knowledge and experience, making it an attractive partner for academic and industry researchers seeking to develop and exploit next generation probes. Probes are a special class of molecules that are administered to patients

Canadian Ebola vaccine enters new trial phase OTTAWA, ON – Canadian Health officials say that the next phase of clinical trials for a promising Ebola vaccine (VSV-EBOV) originally developed by the Public Health Agency of Canada will begin later this fall. As part of this next phase, the vaccine will be tested on volunteers seropositive for human immunodeficiency virus (HIV) infection. The goal of this new clinical trial is to test evidence on the vaccine’s safety and effectiveness at building a protective immune response among HIV-positive people. “It is particularly important to study the effectiveness of this Ebola vaccine in vulnerable populations, such as those living with HIV,” said Dr. Cécile Tremblay, who led the development of the protocol for this study. “These populations can often be most at-risk during outbreaks, because of their compromised immune systems.” The study will build on existing results demonstrating the vaccine’s safety and effectiveness. This includes promising results from the 2014 Phase 1 trials in Canada, a Phase 2 trial on front-line health workers, and a Phase 3 ring-vaccination trial in Guinea during the Ebola outbreak. Initial results from the ring-vaccination trial in Guinea were originally published in The Lancet in July 2015, with final results expected later in fall 2016. The study is to be managed by the Canadian Immunization Research Net-

work (CIRN), with trial sites in Ottawa, Montréal, Senegal and Burkina Faso. Principal investigator Dr. Tremblay (Centre de Recherche du Centre Hospitalier de l’Université de Montreal) and coordinating principal investigator Dr. Joanne Langley (Dalhousie University) will co-lead the Canada-Africa collaborative research team. The Ebola vaccine trial continues to be facilitated and supported by Canada’s International Development Research Centre, Global Affairs Canada, the Canadian Institutes of Health Research, and the Public Health Agency of Canada. The first Canadian vaccinations are anticipated for November 2016, with trials beginning in Africa next year. To see this story online visit http://www.laboratoryfocus.ca/ canadian-ebola-vaccine-enters-newtrial-phase/

in order to visualize their cancers using non-invasive imaging devices like PET scanners. In addition to enhancing the ability to detect the size and shape of cancers, molecular probes can reveal characteristics about the disease that can help select the optimal therapy and also tell physicians very early on if a treatment is working. Rob Baker, McMaster’s vice-president of research, says the collaboration speaks to the reputation and strength of the CPDC, and builds on McMaster’s focus on innovative international partnerships. “This is one of those perfect pairings: blending top science, commercialization and renowned academic and clinical partners where the outputs will benefit patients, the world over,” he says. “We are excited about launching a major collaborative program with one of the premier cancer research institutions,” adds CPDC founder Dr. John Valliant. “The IPDC provides a mechanism to collaborate and create innovative imaging probes that improve cancer care while providing the relationships and mechanisms needed to disseminate new discoveries and help patients on an international scale.” McMaster’s Centre for Probe Development and Commercialization

news (CPDC) is a world leader in the development and translation of new diagnostics, notably molecular imaging probes derived from medical isotopes. The CPDC has extensive experience using cyclotrons and innovative chemical methods to create next generation imaging technologies including companion diagnostics to new therapies. CPDC was created with funding from the Networks of Centres of Excellence (NCE) Centres of Excellence for Commercialization and Research (CECR) program, the Ontario Institute for Cancer Research and several industry partners. From achieving the first remissions in childhood cancer with chemotherapy in 1948, to developing the very latest new therapies, Dana-Farber Cancer Institute is one of the world’s leading centers of cancer research and treatment. It is the only center ranked in the top 4 of U.S. News and World Report’s “Best Hospitals for both adult and pediatric cancer care.” To see this story online visit http://www.laboratoryfocus.ca/cpdcteams-with-dana-farber-cancer-institute-on-cancer-research-initiative/

Food-poisoning bacteria may be behind Crohn’s disease HAMILTON, ON — People who retain a particular bacterium in their gut after a bout of food poisoning may be at an increased risk of developing Crohn’s disease later in life, according to a new study led by researchers at McMaster University. Using a mouse model of Crohn’s disease, the researchers discovered that acute infectious gastroenteritis caused by common foodpoisoning bacteria accelerates the growth of adherent-invasive E. coli (AIEC) – a bacterium that has been linked to the development of Crohn’s. According to the researchers, even after the mice had eliminated the food-poisoning bacteria, researchers still observed increased levels of AIEC in the gut, which led to worsened symptoms over a long period of time. The study, published in the journal PLOS Pathogens, was funded by grants from the Canadian Institutes of Health Research and Crohn’s and Colitis Canada. Crohn’s disease is a debilitating bowel disease characterized by the inflammation of the intestines. Today, one in every 150 Canadians is living with Crohn’s or colitis, a

rate that ranks among the highest worldwide. “This is a lifelong disease that often strikes people in their early years, leading to decades of suffering, an increased risk of colorectal cancer, and an increased risk of premature death,” said Brian Coombes, senior author of the study. At McMaster University he is a professor of biochemistry and biomedical sciences and a researcher at the Michael G. DeGroote Institute for Infectious Disease Research. According to Coombes, the study’s results mean that new diagnostic tools should be developed to identify AIEC-colonized individuals who may be at greater risk for Crohn’s disease following an episode of acute infectious gastroenteritis. “We need to understand the root origins of this disease – and to use this information to invigorate a new pipeline of treatments and preventions. It has never been more pressing.” To see this story online visit http://www.laboratoryfocus.ca/ food-poisoning-bacteria-may-bebehind-crohns-disease/

November/December 2016 Laboratory Focus www.laboratoryfocus.ca

Appointments

Quark Venture Inc., a venture capital investment firm, has hired Karimah Es Sabar as its new CEO while also naming her a partner at the firm. Ms. Es Sabar, most recently led the Centre for Drug Research and Development as president & CEO, and before that,

Karimah Es Sabar

Life Sciences BC. Her expansive global career spans multi-national pharmaceutical/biotechnology companies, start-ups, not-forprofit organizations and venture investment. She has also received multiple awards and recognitions for her pioneering work and holds an Executive Certificate in Management and Leadership from the MIT Sloan School of Management, a MSc degree in Neurochemistry from the Institute of Psychiatry – University of London, England – and a BSc Joint Honours degree in Biochemistry/Chemistry from the University of Salford in Manchester, England. The PEI BioAlliance and Natural Products Canada (NPC) welcome two new faces to NPC’s Charlottetown head office as Karen Wight, VP Investment and Finance, and Stephen Ball, Regional Director of NPC-Atlantic, have joined the organization. Wight and Ball are two of five new hires that will be tasked with fulfilling NPC’s mandate to align and optimize Canada’s assets – natural resources, technologies, platforms, expertise, and world-class research – into a vibrant, pan-Canadian natural products ecosystem. Additional hires across the country include Sue Coueslan, VP Communications and Stakeholder Relations; and two regional direc-

tors: David Gauthier of NPC-West will work closely with the regional node, Ag-WestBio in Saskatoon; and Paul-Thomas Lacroix of NPCQuebec will collaborate with Quebec’s regional node, the Institute of Nutrition and Functional Foods, Université Laval. Other regional directors will be recruited in the near future.

been the director of finance for SolutionInc, and started her career as an audit manager in the assurance and advisory group at PwC. Stephens will continue to serve Immunovaccine as chief financial officer through December 2016 while the company conducts its ongoing process of identifying and transitioning her successor.

Xenon Pharmaceuticals Inc. announces that Dawn Svoronos has been appointed to its board of directors. Ms. Svoronos is a pharmaceutical industry executive whose experience includes a 23-year career at Merck & Co., as well as board positions at Medivation Inc. and Theratechnologies Inc. Svoronos retired in 2011 from Merck & Co., most recently serving as president of Europe and Canada.Previously held positions with Merck include vice president of Asia Pacific and vice president of Global Marketing for the Arthritis, Analgesics and Osteoporosis franchise.

Aquinox Pharmaceuticals has hired Dr. Barbara Troupin as its new chief medical officer and vice president, clinical development. In her new role, Dr. Troupin will lead overall clinical and medical affair strategies for Aquinox’s ongoing programs as well as future development and potential commercialization plans. Dr. Troupin most recently served as senior vice president and chief medical officer at Apricus Bioscience, Inc. where she led the development and execution of clinical strategy for three drug development programs ranging from proof-of-concept to NDA filing. Prior to Apricus, Dr. Troupin held the role of vice president in Medical Affairs at VIVUS, Inc. Dr. Troupin received her Doctorate in Medicine in 1995 from the University of Pennsylvania School of Medicine where she also completed her Master’s in Business Administration, with an emphasis in health care management, from the Wharton School of Business.

Kimberly Stephens has joined Appili Therapeutics Inc. as chief financial officer, bringing with her over 15 years of financial management experience in senior financial positions with private and public companies. Most recently, she

Kimberly Stephens

served as CFO of Immunovaccine Inc. from 2011 to 2016, where she managed the raising of more than $40 million through equity financings and government grants. Prior to joining Immunovaccine Inc., she served as director of finance for the Canadian subsidiary of Germanischer Lloyd. She has also

Alexandra D.J. Mancini has joined InMed Pharmaceuticals, Inc. in the role of senior vice president, Clinical and Regulatory Affairs. Ms. Mancini has over 30 years’ global biopharmaceutical R&D experience with a particular emphasis on clinical development and regulatory affairs. In addition to her activities via her private consulting company True North Synergy, she has been an executive with several biotech companies. As Sr. VP of Clinical & Regulatory Affairs at Sirius Genomics, her role included identifying and managing external resources for medical expertise in sepsis; clinical data management; and statistical theory, programming and analyses. While at INEX Pharmaceuticals as Sr. VP of Clinical & Regulatory Affairs, Ms. Mancini oversaw Clinical Research, Medical Affairs, Clinical Data Management, Medical Writing, Regulatory Affairs, and Quality Assurance

for oncology. She served as VP of Regulatory Affairs at QLT Inc. for oncology and ocular diseases, playing a significant role in the development of VISUDYNE® from the preclinical stage through to its approval as the first drug for agerelated macular degeneration. Ontario Genomics appoints Ihor P. Boszko as its new vice president, Business Development. Boszko will lead and manage all aspects of the organization’s business development activities in

Ihor P. Boszko various sectors including human health, agriculture, mining, water and forestry. For nearly 15 years he has been developing, commercializing and partnering innovative technologies in the medical diagnostics industry. Most recently, in his role as vice president, Business Development of Xagenic Inc., Mr. Boszko led the company’s global commercialization plan for a breakthrough molecular diagnostics platform. Panasonic Healthcare Corp. of North America has hired on Murray Wigmore as vice president of Sales. In his new position, Wigmore will manage a multi-channel distribution and consultative sales team and will be responsible for sales in North America, South America and adjacent markets associated with the life science, pharmaceutical, clinical and industrial laboratory industry. Mr. Wigmore joins Panasonic Healthcare Corporation of North America most recently from Sartorius Japan K.K, where he served as president and managing director.

www.laboratoryfocus.ca

Laboratory Focus November/December 2016

Pharma Notes

NuvoBio Corporation (formerly GeneSpans) (Ottawa, ON) is partnering with Designed Biologics (Ottawa, ON) to develop a new generation of “smart biologic” drugs to treat prostate cancer. Under the terms of the collaboration, NuvoBio will provide funding to help Designed Biologics develop smart biologics, a new class of small peptides (30-75 amino acids) that target specific protein(s) of interest using alternative properties of protein interactions. The funding will support the project through “in-vitro & in-vivo testing” phases. In return, NuvoBio has received the exclusive licensing rights to certain IP and products resulting from the research. Financial terms of the deal were not disclosed. OncoGenex Pharmaceuticals (Vancouver, BC) reports that its Phase 2 Borealis-2™ trial of apatorsen in combination with docetaxel treatment has met its primary endpoint of overall survival. Final analysis from the trial which enrolled 200 patients with metastatic bladder cancer whose disease had progressed following first-line platinum-based chemotherapy showed promising results. Patients who received apatorsen treatment experienced a 20 per cent reduction in risk of death, compared to patients receiving docetaxel alone (HR=.80; 95% CI: 0.65-0.98; p=0.078). The primary analysis was a superiority test of overall survival, performed at a one-sided 0.10 significance level using a stratified log-rank test. The trial was conducted by the Hoosier Cancer Research Network at 28 sites across the U.S. Safety results in patients treated with apatorsen and docetaxel were similar to those observed in patients treated with docetaxel alone. Apatorsen is designed to inhibit production of heat shock protein 27 (Hsp27) to disable cancer cells’ defenses and overcome treatment resistance. Hsp27 is an intracellular protein that protects cancer cells by helping them survive, leading to resistance and more aggressive cancer phenotypes. Sunovion Pharmaceuticals (Marlborough, MA) through its wholly owned subsidiary Sunovion CNS Development Canada ULC has completed its previously an-

nounced acquisition of Cynapsus Therapeutics (Toronto, ON). With the deal complete, Sunovion says it intends to advance the clinical development program of the acquiried formulation of apomorphine (APL-130277), which is a product candidate designed to be a fastacting, easy-to-use, sublingual thin film for the on-demand management of OFF episodes associated with Parkinson’s disease (PD). The acquisition deal values Cynapsus at approximately US$635 million (or approximately CAD$841 million). As a result of the deal, Cynapsus has become a wholly-owned subsidiary of Sunovion and the common shares of Cynapsus will cease to be traded on the NASDAQ Stock Market and Toronto Stock Exchange. The deal was completed by way of a plan of arrangement under the Canada Business Corporations Act. Citing pressures related to pricing, regulatory and market instability, Pharmascience Inc. (Montréal, QC) says it is going into restructuring mode and cutting about 90 positions at its Montreal head office. A generic drugmaker, the company currently ranks as one of the largest pharmaceutical employer’s in Quebec with over 1,500 employees. It makes more than 2,000 generic, over-the-counter and other prescriptions drugs, including 45 million that are filled annually in Canada. Founded in 1983, Pharmascience has experienced constant growth since it was established more than 30 years ago, and has continuously innovated over the years to expand its business and solidify its presence in Canada’s pharmaceutical industry and globally. This is the first major layoff in the company’s history. Diagnostic development firm Zeptometrix Corporation (Buffalo, NY) has filed a statement of claim against Microbix Biosystems (Mississauga, ON) in Canadian Federal Court alleging infringement of its Canadian patent. The claim, (Court File No. T-1672-16), contends that Microbix has violated Zeptometrix’s intellectual property rights and asserts allegations of infringement of 34 claims of its CANADA Patent No. 2,426,172 entitled Nucleic Acid Testing controls. Microbix, a

developer of biological products and technologies, has responded by saying they believe that the allegations are without merit. Zucara Therapeutics (Toronto, ON & Vancouver, BC) has signed a licensing agreement with the Centre for Drug Research and Development (CDRD) for access to a patented set of therapeutic drug compounds that it hopes will help define its lead drug candidate. The two sides are working in partnership to develop unique compounds that they hope will lead to a once-daily therapeutic to prevent hypoglycemia in patients with diabetes. The compounds are pre-clinical, first-in-class antagonists with potential to specifically block somatostatin type 2 receptors, thereby preventing hypoglycemia and restoring a natural glucagon response. Further validation of the lead drug candidate and in vivo studies are ongoing. Zucara says it will be seeking Series A funding in early 2017 to take the company through Phase 1 clinical trials which are expected to start in 2018. Clementia Pharmaceuticals (Montreal, QC) has released top-line results from its Phase 2 clinical trial investigating palovarotene, a retinoic acid receptor gamma agonist (RAR?), for the treatment of fibrodysplasia ossificans progressiva (FOP). FOP is an extremely rare but serious disease in which an accumulation of heterotopic ossification (HO, extraskeletal bone) in muscle and soft tissue progressively restricts movement by locking joints leading to loss of function, physical disability, and risk of early death. Several positive trends were detected in the 40-subject placebocontrolled trial, including palovarotene-related reductions in the proportion of subjects who developed new HO, reductions in volume of new HO, reductions in patientreported pain associated with flareups, and reductions in the time to resolution of FOP-related flare-ups though none reached statistical significance. Additionally, Palovarotene was well-tolerated, with all subjects completing the 12-week trial and enrolling into the open-label extension trial. The full results of the trial are expected to be published next

year. Clementia says it will continue to gather important additional data in the Phase 2 extension trial and in the ongoing observational Natural History Study. Data from these studies will inform the design of a Phase 3 registration trial, which is expected to start in 2017. Antibe Therapeutics Inc. (Toronto, ON) reports that is has completed pre-clinical studies that indicate that its second pipeline drug ATB-352 does not bind to opioid receptors and therefore does not pose a risk of addiction. The company is developing ATB-352 as a pain-killer intended for the relief of acute, severe pain. Currently, opiates, such as oxycontin and fentanyl, are among the most widely used drugs to relieve acute, severe pain, but they are highly addictive. With these promising pre-clinical results, Antibe believes that ATB-352, a novel hydrogen sulfide-releasing analgesic, could offer patients a powerful, safe and non-addictive alternative. ATB352 is a derivative of ketoprofen, a powerful analgesic that carries a high risk of producing gastrointestinal ulcers. However, in animal studies, ATB-352 did not cause ulcers or bleeding, even when given at high doses daily for 5 days. Small amounts of hydrogen sulfide released from ATB-352 protect the lining of the GI tract, and provide additional analgesic and antiinflammatory effects. ProMetic Life Sciences Inc. (Laval, QC) announces that its Phase 2 clinical trial using PBI-4050 to treat patients with metabolic syndrome and type 2 diabetes has been completed, meeting both its primary and secondary endpoints. In addition to safety and tolerability, the open label study was designed to evaluate PBI-4050’s effect on metabolic syndrome parameters as well as on pro-inflammatory/ fibrotic and diabetic biomarkers in blood and urine. PBI-4050 (800 mg) was administered once daily to 24 patients for a period of 12 weeks. Ten of these patients were enrolled in an additional 12 weeks extension. Results show that PBI-4050 was well tolerated with no serious drug related adverse events.

November/December 2016 Laboratory Focus www.laboratoryfocus.ca

feature

b y Sika nd er Gill, Ma xim Slyadnev, and Alexander Strog anov, Lu mex Inst rument s Canada Missio n, BC, Canada

Optimization of paramagnetic bead based nucleic acid isolation for high throughput potato pathogen detection using

Microchip Real-time PCR Introduction The recently developed microchip based real-time PCR offers accurate, robust and cost effective detection of pathogens in potato seed, crop, and soil which is essential for diagnosis, and surveillance for this important food source in addition to various other applications.1-3 The innovative features of this technology impart an edge over the currently used methods for potato pathogen detection as cost effective technology with high throughput screening capability (Table 1). To meet its throughput, a simple, and fast nucleic acid isolation from potato tubers, seeds, crops, and soil with amenability to automation is also essential as an upstream method.4 To address this requirement, a variety of nucleic acid extraction methods are available along with commercial kits.5 Many of these methods involve time-consuming and labour-intensive complex steps such as centrifugation, precipitation, and filtration. The choice of method has a profound impact on the time and effort required to process the screening of potato pathogens. The choice of method is also important when many samples need to be processed. Thus the existing methods are not always suitable while an innovative sample preparation can enhance chances of pathogen detection and imparts ease and speed to the detection process. This is especially important when lab staff has limited training and time. Nucleic acid isolation and purification consists of three basic steps: lysis, removal of protein and contaminants, and recovery of DNA or RNA. Currently, commercial kits using a combination of solvents and solidphase support including paramagnetic beads, and spin columns are widely used to isolate and purify DNA and RNA.6 The paramagnetic beads are especially very popular by providing fast, efficient purification, and carrying pre-concentration of the isolated nucleic acid typically carried out in the physiological range.7-8 The paramagnetic DNA purification is an improvement upon centrifuge-dependent isolation techniques. The need to move the samples into and out of a centrifuge is inefficient for meet-

Table 1 Features and advantages of microchip real-time PCR Feature

Advantages

Miniaturized PCR wells

• Low sample and reagent consumption o 0.5-1.8 µl of 2x master mix o 0.5-1.8 µl sample • Imparts low running cost to screening projects

High thermal cycling rate (10-12°C/s)

• Fast analysis 45 cycles in 20 minutes • Addresses high through screening needs

Low detection limits

• High sensitivity • Can detect 1-5 copies per micro-reactor • Supports early detection of pathogenicity • Offers advantages over conventional techniques used for screening

Microchip with ready to use lyophilize PCR mix

• Reduces labor requirements for screening • Minimizes human error in economically important screening projects • Adds ease and simplification to the screening process • Requires just addition of sample to the microchip • Allows entire panel of relevant pathogens to be detected for each sample

Simultaneous detection in all the micro-reactors of the microchip

• Offers uniformity of data • Displays better comparatives of pathogenicity

Two detector channels-FAM/ SYBR Green and ROX

• Two-plex in each micro-reactor means 60 targets in 30 reactor microchip. • Minimizes complexity of analysis for being only two channels • Minimize manufacturing cost and price to address affordability

Microchip isolated from environment

• Minimized contamination • Adds to reliability of analysis

Chemically modified microchip

• The hydrophobic surfaces of the microchip except the hydrophilic micro-reactor surfaces ensure containment of the sample-mix into the micro-reactors • Minimize cross contamination • Offer reliability by minimizing false positive/negatives

Small footprint (W X D X H) in cm: 25 x 30 x 19

Minimize lab space requirements

Weight: 5 kg

Easy to move on the bench

ing high throughput demands. On the other hand, the use of paramagnetic beads being amenable to automation, allows the tubes to remain on its deck throughout the entire process. In the present study, we have adapted the paramagnetic bead approach to potato samples by isolating the plasmid DNA spiked into the po-

tato sample by replacing silica beads with paramagnetic beads and analysing the isolated DNA by microchip real-time PCR.

Materials and methods 1. Procurement of materials: The silica bead based DNAsorb nucleic acid extraction kit was procured

from AmpliSens, Bratislava, Slovak Republic and paramagnetic beads were purchased from Qiagen, Valencia, CA, USA. Recombinant plasmid pUC-35S gene of cauliflower mosaic virus (CAM virus) was custom synthesised from ATG Service-Gen, St.-Petersburg, Russia.

www.laboratoryfocus.ca

Laboratory Focus November/December 2016

Feature

2. Potato sampling and processing: Sample piece was scooped out from the potato eye using a tiny spatula and placed into the cup of the squeezer ensuring the potato skin to squeeze first (Figure 1). The potato mesh was collected further into a 30 ml falcon tube and was further crushed with spatula followed by weighing out 30 mg in individual 1.5 ml taper tubes for further process.

Table 2 DNA extraction and purification protocol steps for sorbent beads and magnetic beads Feature

Silica beads Applicable Duration (sec)

Paramagnetic beads Applicable Duration (sec)

• Lysis buffer 300 µl • Vortex • Incubation 65°C

Centrifugation 5,000×g

120, 300

+ 300

300

Removal of supernate Addition of sorbent bead 25 µl and mixing Addition of paramagnetic bead addition 15 µl & mixing

120, 300

Centrifugation 5,000×g

Magnet incubation & mixing • Wash solution-I 300 µl • Vortex

Centrifugation 10,000×g

Magnet incubation • Wash solution-II 400 µl • Vortex

Centrifugation 10,000×g

Magnet incubation • Wash solution-II 400 µl • Vortex

Centrifugation 10,000×g

Magnet incubation

Pre-drying 65°C

600

120

• Elution TE buffer 50 µl • Vortex • Incubation 65°C

300

Centrifugation 12,000×g

Magnet incubation

Total duration (except addition and removal of liquids)

~30 min

10 ~25 min

3. Spiking of the potato samples: To each tube containing 30 mg of the sample, 1µl (10 pg/µl) of plasmid pUC-35S was spiked using programmable automatic pipettor (Sartorius AG, Gottingen, Germany). It was followed by addition of 300 µl of lysis buffer of the Ribo-sorb kit and vortexed for 5 minutes. The tubes were then heated at 65°C for 5 minutes using dry bath heater (MK-20, China) with occasional manual mixing. The tubes were centrifuge for 5 sec at 5000×g to remove drops from internal surface of the lids. 4. Addition of mobile solid phase: To one of the tubes, 25 µl of silica beads suspension, and to the other tube 10 µl of paramagnetic beads suspension was added, respectively. The tubes were vortexed for 10 sec and incubated at room temperature for 60 sec and again mixed for 10 sec and incubated for 5 min at room temperature. 5. Separation of beads: The tubes containing silica beads were centrifuge for 30 sec at 10,000×g to pellet the beads for removing and discarding supernate without disturbing the bead pellet. The tubes containing paramagnetic beads were inserted into the magnet block for 1 minute to pellet the beads and remove the supernate. 6. Washing of beads: To each tube, 400 µl of Washing Solution 1 was added and the silica bead containing tubes were vigorously vortexed to fully resuspend the beads followed by centrifugation for 30 sec at 10,000×g. The paramagnetic bead containing tubes were subjected to magnet block for pelleting beads and removal of supernate. To each of the bead containing tubes, 500 µl

November/December 2016 Laboratory Focus www.laboratoryfocus.ca

feature Figure 1 (a) Potato sampling, (b) Microchip real-time PCR analyzer AriaDNA, (c) Microchip

b a

of Washing Solution 2 was added followed by vigorous vortex. The silica bead tubes were centrifuge for 30 sec at 10,000×g. The supernate was carefully removed and discarded. The tubes containing paramagnetic beads were inserted into the magnet block for 1 minutes to pellet the beads and remove the supernate. The procedure was repeated with this buffer. 7. Pre-drying of DNA bound beads: All the tubes were incubated with open caps for 12 min at 60°C for pre-drying in the heater ensuring no over-drying of beads. 8. Elution: The bead pellets were re-suspended in 50 µl of the TE buffer and incubated for 3 min at 60°C with intermittent mixing. 9. Optimization studies: The sample prep was also attempted for using isopropanol for washing the paramagnetic beads during the nucleic acid isolation process. 10. Microchip real-time PCR: a. The real-time PCR was set up for 35S gene of CAM virus. Master mix was prepared following the Table 1, and then 1.2 µl was dispensed into each of the well of the 6x5 format microchip as per the microchip map presented in Figure 1d. The microchip was filled with 620 µl of silicone oil that covered the dispensed test mix in wells to contain evaporation during thermocycling. b. The amplification was performed with thermal condition: Hold 95°C for 120 sec followed by 45 cycles of 95°C for

6 SB-PP

SB-PP

PCR-C

PB-PP

1 sec and 60°C for 30 sec. For each well, a fluorescent light emission was recorded. Analysis was based on the Ct values of the PCR products. The Ct value is the PCR cycle at which the fluorescence measured between each cycle exceeds a threshold determined by background fluorescence at baseline and is placed in the exponential phase of the amplification curve. The threshold and baseline were set automatically by software or were user-defined.

(d) Microchip map, SB-PP (Silica beads and potato spiked with plasmid DNA), PB-PP (Paramagnetic beads and potato with spiked plasmid), and PCR-C (PCR control with the plasmid DNA).

Figure 2

Amplification curves of isolated recombinant plasmid DNA with silica beads (blue curve) and paramagnetic beads (green curve) with a PCR control (red curve).

Figure 3

Comparison of Ct values of both the silica beads (vertical bars =A1-6, and D1-6), paramagnetic beads (horizontal bars=B1-6), and PCR control (black bars=C1-6).

Results & discussion Numerous detection methodologies for pathogens are now available, but their acceptance to perform high throughput screening, faces an important question on sensitivity, accuracy, robustness, frequency of testing, and the cost. Recently, a microchip based fast and simple polymerase chain reaction method has been developed for detection of pathogens in diverse biological and soil samples using miniaturized volumes of samples and reagents.1-3 The features of this technology detailed in Table 1, address these questions. The low running cost of this approach, makes it especially attractive for large-scale screening of crops, poultry, cattle, fish, and food for pathogen detection. With the objective of providing an easy, fast and low cost sample prep method for isolation of bacterial, fungal, and viral pathogen nucleic acid to be used in the microchip, a paramagnetic beads based method is modified and optimized in the present studies. Comparison of silica beads and paramagnetic beads: From 30 mg of potato tissues spiked with 1 µl of

30 Threshhold Cycle (#)

Numerous detection methodologies for pathogens are now available, but their acceptance to perform high throughput screening, faces an important question on sensitivity, accuracy, robustness, frequency of testing, and the cost.

25 20 15 10 5 0

3 4 Microchip well (#)

www.laboratoryfocus.ca

Laboratory Focus November/December 2016

Feature Figure 4

In view of the importance of false negatives in the screening process, it is better to provide larger surface area to enhance the probability of catching pathogen nucleic acid, 20 µl of the beads should be used in this screen.

Optimization of quantity of paramagnetic beads (one amplification curve from five replicates (n=5) is presented for better display. The green, red, sky blue, violet, and brown curves are for 5, 10, 15, and 20 µl of paramagnetic beads, respectively. All the set were run with 30 mg of potato tissue except no potato tissue for blue curve. The yellow curve is run with 1 µl of the plasmid as a PCR control.

Figure 5

Simplification of wash process using isopropanol. The violet curve represents the sample prep using isopropanol and the yellow curve PCR control with 1 µl of the plasmid (n=5).

plasmid DNA, the DNA extracted using paramagnetic beads resulted amplification Ct values comparable to silica beads which are supplied as optimized reagent by the kit provider (Figure 2). The n=12 replicates of both the paramagnetic beads, and silica beads, were found to be very close to one another (Figure 3). Optimization of paramagnetic beads: With the objective of optimizing the quantity of paramagnetic beads, a set of extractions were carried with 30 mg of potato spiked with 1 µl of plasmid DNA with addition of 5, 10, 15, and 20 µl of paramagnetic bead suspension. The results indicated that 20 µl of the beads was slightly better than the other samples even

though there was not a significant difference among them (Table 4). In view of the importance of false negatives in the screening process, it is better to provide larger surface area to enhance the probability of catching pathogen nucleic acid, 20 µl of the beads should be used in this screen. The data also suggest that DNA extraction with both silica beads and paramagnetic beads resulted similar quantity of DNA in the presence and absence of 30 mg of potato tissue. The extraction efficiency: The efficiency of DNA extraction does not seem affected by the 30 mg of potato tissue used in the extraction process. This observation is supported by the Ct values of the DNA extracted from

the control (1 µl in 30 µl of TE buffer) using 10 µl of paramagnetic beads which is close to that of the 30 mg of potato extracted with 10 µl of the beads (Figure 4). 3. Optimization of wash process of paramagnetic beads: The sample prep was also attempted with isopropanol for washing the paramagnetic beads during the nucleic acid isolation process. The results indicated that two washes of 200 µl isopropanol were equivalent to the standard wash solution of the kit (Figure 5). This suggests that sample prep can be simplified with isopropanol which in turn reduced the pre-drying step from 5-10 min to 1-2 min (n=5). The number of the ct values (29.7) for the extraction with isopropanol corresponds with that of the standard wash buffers used in the previous experiment while the control displayed the ct 20.46. Total duration of protocols: The duration of each step has been presented in the Table 2 where addition and removal of liquid is not included for easy comparison of total duration of both sorbent beads and paramagnetic beads. The latter technique is about 5 minutes shorter, easier to perform using magnet block rather than centrifugation, and also allows to avoid inconsistent removal of supernate due to pellet disturbance. Moreover, the use of paramagnetic beads is amenable to automation as magnet block can be used on the deck of automated workstations for high throughput isolations.4

References: 1. Tanaka, Y., Slyadnev, M.N., Sato, K., Kitamori. K.: Acceleration of an Enzymatic Reaction in a Microchip. Analytical Sciences. 2001,17: 809-810 2. Navolotskii D.V., Perchik A.V., Mark’yanov I.A., Ganeev A.A., and Slyadnev M.N.: Microchip Analytic System for Multiplex

Analysis by Real-time Polymerase Chain Reaction with Reagents Immobilized in Microreactors. Applied Biochemistry and Microbiology, 2011, 47: 221–227 Applications of AriaDNA: http:// www.lumexinstruments.com/ catalog/pcr_analysis/aria_dna. php#buklet Gill, S., Gill, R., Lemay, M., and Liand, D.: Validation of MO BIO’s PowerMagTM Soil DNA Isolation Kit on the Automated Liquid Handling Workstation. https:// mobio.com/media/wysiwyg/ Poster-Automation%20of%20 Mo%20Bio%20PowerMag %20 Soil%20DNA%20Kit%20on%20 VERSA%201100%20Workstation%20[Compatibility%20Mode]. pdf Anandika D.: DNA Extraction and Purification. Materials and Methods. 2013, 3: 191 Tan S.C., Yiap B.C.: DNA, RNA, and Protein Extraction: The Past and The Present. J Biomed Biotechnol. 2009; article id 574398. Saiyed, Z.M., Bochiwal, C., Gorasia, H., Telang, S.D., Ramchand, C.N.: Application of magnetic particles (Fe3O4) for isolation of genomic DNA from mammalian C.N. cells. Analytical Biochemistry. 2006, 356: 306-308. Azimi, S.M., Nixon,G., Ahern, J., Balachandra, W.: A magnetic bead-based DNA extraction and purification microfluidic device. Microfluidics and Nanofluidics. 2011, 11: 157–165.

To see this story online visit http://laboratory focus.ca/optimization-ofparamagnetic-bead-basednucleic-acid-isolation-for-highthroughput-potato-pathogendetection-using-microchipreal-time-pcr/

November/December 2016 Laboratory Focus www.laboratoryfocus.ca

feature

b y Bria n Riv era , Prod uct Ma na ger , Biose parations & Spe cial ty LC, Phe nom e ne x

Orthogonal Methods and Mobile Phase Optimization for Liquid Chromatography Analysis of Modified Synthetic Oligonucleotides Table 1 OGNs Sequences and Modifications Introduction Synthetic oligonucleotides (OGNs) are becoming increasingly popular as novel agents for the treatment of disease. The characterization of OGNs is important in the drug development process, and one common method is liquid chromatography, with detection either by UV at 260 nm or coupled with mass spectrometry or tandem mass spectrometry (LC–MS or LC–MS/MS). When characterization of the OGN is not needed, strong anion-exchange HPLC (SAX-HPLC) can be used, because this high-resolution technique can be used to separate n-1 failure sequences. Although analysis of so-called “vanilla” or unmodified OGNs can be relatively straightforward, other modified OGNs present challenges. Several different modifications to the nucleic acid can be made to prevent nuclease degradation; these include backbone (e.g. phosphorothioate), ribose/ deoxy-ribose sugar (e.g. bridged nucleic acid), and 2’-OH substitutions. Additionally, various different conjugations can be performed to either the 5’ or 3’ end of the oligo. Here we investigate chromatographic separations for several OGNs with various modifications. Depending on the separation mode used, method parameters such as mobile phase and gradient program were adjusted to optimize separation – specifically, separation of the n-1 failure sequence.

Materials and Methods Crude, desalted OGNs (Bridged Nucleic Acid, Amino C12, DNA Phosphorothioate, and 2’-MOE Gapmer) were purchased from Integrated DNA Technologies (Coralville, IA, USA). Sequences, along with modifications, are indicated in Table 1. These specific OGNs were selected to represent the various modifications used to improve pharmacokinetic profile and increase nuclease stability. Physiochemically, they can increase electrostatic interactions and/or add hydrophobicity to the molecule, and these two character-

Oligo Type

Length

Sequence

Bridged Nucleic Acid

19mer

5’-TA/BNA-A/TA/BNA-meC/GT/BNA-T/TA/BNA-T/AC/BNA-G/CC/BNA-C/A-3’

Amino C12

24mer

5’AmMC125-TCGTGCTTTTGTTGTTTTCGCGTT-3

DNA Phosphorothioate

19mer

5’-ACT*G*A*C*T*G*A*C*T*G*A*C*G*T*A*C*T-3

2’-MOE Gapmer

20mer

5’- mC mG mA mC mU A T A C G C G C A A mU mA mU mG mG -3’

istics will determine the respective OGN behavior by reversed phase and ion exchange chromatography.

OGN Analysis by Ion Pair Reversed Phase OGNs, which are relatively polar due to a ribose or deoxyribose sugar and phosphodiester backbone, need to be analyzed by ion pair reversed phase. Typically, alkyl amines such as triethylamine are used. Triethylammonium acetate is a commonly used mobile phase for LC-UV assays, however, TEA-Hexafluoroisopropanol (HFIP) is more common in LC-MS. HFIP not only helps with electrospray desporption1 but also helps with peak shape and overall chromatographic performance.2,3 Recently, other alkylamine ion pairing agents have been used as well. This includes N-diisopropylethyl amine (DIEA), which has been reported to also improve electrospray desorption. Additionally, the utilization of different alkylamine ion pairs can have profound effects on chromatography – specifically improvements in peak shape and separation of n-1 failure sequence. Figures 1 and 2 show performance using the more commonly used TEA compared to DIEA. Note the improvement in both peak shape as well as resolution of the n-1 failure sequence.

OGN Analysis by Ion Exchange Chromatography Ion exchange is commonly used for OGN characterization because of its high resolution and relatively easy

Figure 1

Chromatogram for BNA, DIEA-HFIP Mobile Phase (Phenomenex App ID 23656)

Method 1: DIEA and HFIP LC Column: Clarity 2.6 µm Oligo-XT Flow Rate: 0.3 mL/min Dimensions: 50 x 2.1 mm Temperature: 60° Part No.: 00B-4746-AN LC System: Agilent 1200 Mobile Phase: A: 50 mM HFIP & 5 mM DIEA in Water, B: 50 mM HFIP & 5mM DIEA in Acetonitrile Detection: UV-Vis @ 260 nm Gradient: 5 – 20% B in 15 min to 95% wash for 5 min

Figure 2

Chromatogram for BNA, TEA-HFIP Mobile Phase (Phenomenex App ID 23662)

Method 2: TEA and HFIP LC Column: Clarity 2.6 µm Oligo-XT Flow Rate: 0.3 mL/min Dimensions: 50 x 2.1 mm Temperature: 60° Part No.: 00B-4746-AN LC System: Agilent 1200 Mobile Phase: A: 100 mM HFIP & 4 mM TEA in Water, B: 100 mM HFIP & 4 mM TEA in Methanol Detection: UV-Vis @ 260 nm Gradient: 10 – 25% B in 15 min to 95% wash for 5 min

www.laboratoryfocus.ca

Figure 3

Amino C12 Linker DNA (Phenomenex App ID 23829) Ion Exchange Chromatography Method for Amino C12 Linker LC Column: Clarity 5 µm Oligo-SAX Flow Rate: 1.6 mL/min Dimensions: 250 x 4.6 mm Temperature: 30° Part No.: 00G-4749-E0 LC System: Agilent 1200 Mobile Phase: A: 20 mM Tris Base, pH 8.0, B: 20 mM Tris + 1.25 M NaCl,pH 8.0 Detection: UV-Vis @ 260 nm Gradient Program: 32 -46.6% in 10 min

Figure 5

feature

BNA 19mer (Phenomenex App ID 23831) Ion Exchange Chromatography Method for BNA LC Column: Clarity 5 µm Oligo-SAX Flow Rate: 1.6 mL/min Dimensions: 250 x 4.6 mm Temperature: 30° Part No.: 00G-4749-E0 LC System: Agilent 1200 Mobile Phase: A: 20 mM Tris Base, pH 8.0, B: 20 mM Tris + 1.0 M NaOCl4pH 8.0 Detection: UV-Vis @ 260 nm Gradient Profile: 10-46.6% in 10 min

Figure 4

Laboratory Focus November/December 2016

DNA 19mer Phosphorothioate by Ion Exchange (Phenomenex App ID 23833)

method implementation. This is especially true when unmodified OGNs are being analyzed. It is often preferred for quality control applications such as impurity assays where characterization by MS is not required. One issue that may arise is optimization of mobile phase when analyzing an OGN that has a modification that specifically increases binding affinity – that is, an OGN that exhibits higher electrostatic effects. OGNs with ribose modifications in particular can be somewhat problematic. An example would be the Bridged Nucleic Acid (BNA) modifications, which have a higher binding efficiency and thus exhibit higher electrostatic interactions.4 Although chloride is used for many methods, strongly adsorbing OGNs like BNAs may require a higher ionic elution strength such as perchlorate provides. Figure 3 shows a good separation of n-1 failure sequence using the perchlorate mobile phase. One challenging modification to the OGN when working with ion exchange are conjugations to lipophilic tails. Because the non-porous particle used with ion exchange resins is inherently hydrophobic, it must be coated with a hydrophilic layer to ensure no secondary interactions cause peak broadening. As an example, the Amino-C12 linker conjugate was used as a surrogate for an OGN linked to a lipophilic compound. As we see in Figure 4, the Amino-C12 linker DNA shows good peak shape, whereas a more hydrophobic phase might show slight peak broadening. Because the OGN has an unmodified phosphodiester backbone, a typical mobile phase using sodium chloride was implemented.

Ion Exchange and IP Reversed Phase as Orthogonal Methods

Figure 6

Chromatogram for DNA Phosphorothioate by Ion Pair Reversed Phase (Phenomenex App ID 23657)

One other consideration with ion exchange methods is working with phosphorothioates. This modification yields diastereomers, which, when analyzed by ion exchange, will show a partial separation.5 However, with multiple phosphorothioate modifications, which many therapeutic platforms will utilize, the number of diastereomers increases significantly and excessive peak broadening may occur. Figure 5 shows a primary example of this, where the n-1 impurity only shows a partial shouldering. In this case, it might be preferable to run an orthogonal ion pair reversed phase method to confirm the presence of the impurity, as seen in

Figure 6. This can be used even if the intent is not LC-MS/MS but simply separation of the n-1 impurity (i.e. an impurity assay) by LC-UV.

Summary With various modifications for OGNs, several considerations should be taken into account when developing an analytical method. This is dependent on the intent for the analysis (e.g. full characterization by LC-MS/MS, impurity profile, etc.) as well as limitations in the separation mode. Here we showed two chromatographic separation modes, ion pair reversed phase and ion exchange. Depending on the modification to the OGN, adjustments to each method should be made, in addition to the utilization of both in cases where an orthogonal separation is needed.

References 1. Rudge, James et al. “Preparation and LC/MS Analysis of Oligonucleotide Therapeutics from Biological Matrices.” Chromatography Today. 1 Mar. 2011: 16-20. Print. 2. Bonilla, Jose. “Handbook of Analysis of Oligonucleotides and Related Products.” 2011. 10-11. Print. 3. Sun, Hongguang, Xun Zhu, Patrick Y. Lu, Roberto R. Rosato, Wen Tan, and Youli Zu. “Oligonucleotide Aptamers: New Tools for Targeted Cancer Therapy.” Molecular Therapy—Nucleic Acids Mol Ther Nucleic Acids 3.8 (2014): n. pag. Print. 4. Obika, Satoshi, Osamu Nakagawa, Akiko Hiroto, Yoshiyuki Hari, and Takeshi Imanishi. “Synthesis and Properties of a Novel Bridged Nucleic Acid with a P3’ N5’ Phosphoramidate Linkage, 5’-amino2’,4’-BNA.” Chem. Commun. 17 (2003): 2202-203. Print. 5. Frederiksen JK, Piccirilli JA. Separation of RNA phosphorothioate oligonucleotides by HPLC. Methods Enzymol. (2009). 468:289309. Print.

To see this story online visit http://laboratory focus.ca/orthogonal-methodsand-mobile-phaseoptimization-for-liquidchromatography-analysis-ofmodified-syntheticoligonucleotides/

November/December 2016 Laboratory Focus www.laboratoryfocus.ca

Feature

b y Natha lie Chandelier , Vincent Du fey, Mu riel Art, Ep p end o rf Appl ication Te chnol ogie s S.A ., Nam ur, Be l gium Dia na Hü bler , Eppe ndorf Ag, Ham bur g, Ge r m any

Fast, Precise and Sterile Liquid Transfer in Cell Culture with

Multipette® M4 /Repeater® M4 Abstract The most important factors in cell culture are sterility, accuracy, and precision. Positive displacement dispensers offer the great advantage of complete aerosol prevention as the piston integrated in the tip of the positive displacement system hermetically seals the sample in the tip to prevent contamination. In regard to accuracy and precision positive displacement dispensers show equivalent results to those obtained with air-cushion pipettes. In contrast, reproducibility of results will be affected when pipette controllers in combination with serological pipets are used.

Introduction Animal cell culture has become a common laboratory technique and is used for a large range of applications. The most important factors to consider are sterility in terms of contamination prevention and accuracy, as well as precision. Commonly, pipette controllers with serological pipets are used for the transfer of large volumes. Air-cushion pipettes are used for all other parts of the application. But one of the greatest sources of contamination is aerosol formation during pipetting, therefore using a two-phase filter tip is recommended by Eppendorf. An alternative to these systems are positive displacement dispensers, such as the Multipette M4 (U.S.: Repeater M4), that prevent contamination of the instrument or cross-contamination via aerosols completely. This is ensured by a piston that hermetically seals the sample in the tip of the positive displacement system. Furthermore, this system allows for accurate and precise pipetting results especially when working with liquids whose physical properties differ from those of water (e.g. DMSO, cell culture medium).

Materials and methods Instrument calibration Systematic and random errors were determined by gravimetric method in accordance with EN ISO 8655:2002.1

Table 1

Figure 1 and 2

Cell counting results obtained with three different pipetting systems used to dispense 100 mL of cell solution (n=30)

Systematic (Fig.1) and random error (Fig.2) at 100 mL measured for the different pipetting systems used

www.laboratoryfocus.ca

Laboratory Focus November/December 2016

feature Figure 3

The results show that using a positive displacement system, such as the Multipette M4/Repeater M4 in combination with Combitips advanced, for cell culture applications, ensures reproducibility of the final results. The volumes tested were 100 μL in each pipetting system. Cell counting Cell number was determined by using the CASY® Cell Counter and Analyser, model TT 150 μm (Roche®, Basle, Switzerland). Measurement was performed by suspending 100 μL of cell suspension in 10 mL of CASY ton.

Figure 4

Cell seeding and WST-1 colorimetric assay After cell counting, a cell suspension of 1.5 x 106 HEK 293 cells/mL was prepared for cell seeding. Various quantities of cells were seeded into 24-well plates (Eppendorf). 40 μL of Water Soluble Tetrazolium salt (WST-1; Roche) were added per well. Cell culture plates were incubated for 3 h (37 °C, 5 per cent CO2). The plates were mixed in a plate reader and read at 450 nm (reference: 690 nm).

Results and discussion

Figure 5

Chromatogram for BNA, DIEA-HFIP Mobile Phase (Phenomenex App ID 23656)

Figure. 3 – 5: Signal curves generated by increasing numbers of cells. Three different pipetting systems were used for cell seeding: Xplorer (Fig. 3), Multipette M4/Repeater M4 (Fig. 4), and Easypet 3 (Fig. 5).

Performance evaluation of the used pipetting systems Each instrument was used with its dedicated consumable (Multipette M4 /Repeater M4 with Combitips advanced® 1 mL, Eppendorf Research® plus pipette with ep Dualfilter T.I.P.S.® 100 μL, Eppendorf Xplorer® pipette with ep Dualfilter T.I.P.S. 1 mL, and Easypet® 3 with 1 mL Eppendorf Serological Pipets). Easypet 3 is the least accurate and least reproducible system of all four systems (Figures 1 and 2). In contrast, with mean systematic and random errors below 0.6 %, aircushion pipettes and Multipette M4/ Repeater M4 perform as the most reliable systems. Impact of the pipetting system on cell counting To evaluate the impact of the three pipetting systems on cell counting a manual air-cushion pipette (Eppendorf Research plus) with ep Dualfilter T.I.P.S., a positive displacement system (Multipette M4/Repeater M4) with Combitips advanced, and an electronic pipette controller (Easypet 3) with serological pipettes were compared. The number of HEK 293 cells was determined using the CASY technology.

As shown in Table 1, the level of cell viability is around 96 per cent for all conditions assessed. The cell count variability of the positive displacement dispenser and the air-cushion pipette is almost equivalent while the reproducibility obtained with the pipette controller is significantly lower. Impact of the pipetting system on cell seeding Based on the cell counting data, six cell suspensions of 1.5 x 106 cells/mL were seeded into 24-well plates. The electronic Xplorer pipette with ep Dualfilter T.I.P.S., the Multipette M4/Repeater M4 with Combitips advanced, and the Easypet 3 with serological pipettes were used for cell transfer. 40 μL/ well of WST-1 were added. WST1 is cleaved to formazan by cellular enzymes largely dependent on the NAD(P)H production in viable cells and therefore directly correlates to their number (Figure 3 to 5).

Conclusion The results show that using a positive displacement system, such as the Multipette M4/Repeater M4 in combination with Combitips advanced, for cell culture applications, ensures reproducibility of the final results. Data obtained with the dispenser are less variable than those obtained with aircushion pipettes. Reproducibility of the final results will be affected when a less precise instrument such as a pipette controller is used. Combining accuracy, precision, and contamination prevention, the Multipette M4/Repeater M4 in combination with Combitips advanced represents a perfect alternative for all routine tasks requiring a liquid handling device in cell biology labs.

References 1. EN ISO 8655:2002, Parts 1– 6. Piston-operated volumetric apparatus. Beuth-Verlag, Berlin, Germany.

To see this story online visit http://laboratory focus.ca/fast-precise-andsterile-liquid-transfer-in-cellculture/

November/December 2016 Laboratory Focus www.laboratoryfocus.ca

New Products MICROSCOPES The ZEISS Axio Observer family consists of three stable and modular microscope stands for flexible and efficient imaging. Among its features is Autocorr, a technology that allows adaption of immersion objectives to varying sample carriers and imaging conditions. The correction of spherical aberrations results in improved image contrast, higher resolution and more efficient fluorescence detection. The new Autoimmersion option that builds up and maintains stable water immersion without traditional problems such as evaporation or air bubbles is compatible with multiple water immersion objectives in the nose piece. Definite Focus.2 is a novel hardware focus that automatically compensates for focus drift and enables long-term time-lapse imaging and demanding multi-position experiments. The ZEISS Axio Observer can be combined with the new multicolor LED light source Colibri 7 that delivers up to seven excitation wavelengths. In combination with the new Virtual Filter technology users can perform ultrafast fluorescence imaging with high spectral flexibility. Among its other features are wealth of interfaces for imaging technologies, ranging from widefield transmitted light to 3D sectioning with Apotome.2, fast and sensitive confocal superresolution imaging with ZEISS LSM 880 and Airyscan, and structured illumination as well as photoactivated localization microscopy with Elyra PS.1.

Web: www.zeiss.com/axioobserver

FUMES & HOODS The new Air Science Safefume™ Cyanoacrylate Fuming Chamber is designed to safely develop latent fingerprints using ethyl cyanoacrylate (CNA) vapour in controlled environments. The chamber’s stamper resistant compartment ensures chain of custody is maintained, while its unique Air Science Multiplex™ filtration system, together with professional design and unique construction features offers personnel protection during use. Its ductless filtration system requires no connection to an outside exhaust system, while its automatic control system programs the fuming cycle. A versatile system of hanging rods and shelves allows proper positioning of items of evidence in the chamber. The Safefume is available in 24”, 30”, 48”, 60” and 72” models.

Web: www.airscience.com/safefume-fuming-chambers

PIPETTES BrandTech introduces the Transferpette® S Adjustable Volume, single channel pipette, now in 0.1-2.5µL and 5-50µL sizes. Constructed with innovative composite materials, Transferpette® S pipettes are lightweight and fully autoclavable, and accept tips from most manufacturers. True one-handed selection, along with light weight and short pipetting stroke reduce the risk of repetitive strain injury. Its 4-digit display is easy to read for precise volume settings, and is always visible during pipetting.

Web: www.BrandTech.com

DNA SEQUENCING Fluidigm Corporation’s Juno™ Targeted DNA Sequencing Library Preparation System is a scalable automated library preparation solution for medium-to-high-throughput next-generation sequencing (NGS). The system brings the Fluidigm® Juno microfluidic automation platform together with the benefits of multiplexed target amplification allowing laboratories to accurately sequence more samples faster. Optimized for use with Illumina sequencers, the Juno Targeted DNA Sequencing Library Preparation System also redefines the library preparation workflow from extracted DNA to NGS-ready amplicon libraries. Supporting a range of panel designs from targeted multiplexed panels covering a few user defined genes to more comprehensive panels of up to 4,800 amplicons from hundreds of genes, the system can help laboratories achieve new sequencing efficiencies with consistent performance.

Web: www.fluidigm.com

SOFTWARE Bio-Rad Laboratories, Inc. announces the release of its KnowItAll® 2017 spectroscopy software and databases. The new version offers an additional 976,000 reference spectra in the KnowItAll Spectral Library including Bio-Rad’s Sadtler™ spectra as well as spectra from John Wiley & Sons, Inc., bringing the spectral database collection to over 2.3 million spectra. With the update, users can save time by automatically importing structures and properties directly from PubChem to enhance user-built databases. The improvements also help to streamline Bio-Rad’s Enterprise Server software for management of spectral and chemical data throughout an organization. Bio-Rad’s KnowItAll® ID Expert software application for spectral identification also supports all techniques in the KnowItAll software (Infrared (IR), Raman, NIR, NMR, MS, UV-Vis).

Web: www.knowitall.com/whatsnew

MIXERS MilliporeSigma’s Mobius® Power MIX 2000 single-use mixing system is engineered with advanced technology to effectively handle difficult-to-mix buffers, culture media powders and other pharmaceutical ingredients. The Mobius® Power MIX achieves a strong vortex using an impeller design and motor based on magnetically-coupled NovAseptic® technology, a proven mixing technology in stainless steel tanks. The system provides accessible, sterile zero deadleg sampling directly from the mixing container. A probe port allows for insertion of either a reusable probe for non-aseptic processes or a pre-sterilized, single-use sensor for in-process pH measurement of aseptic processes. Other design features make the mixing system easy to use, enabling loading of bags by one person and offering an optional integrated hoist for powder delivery.

Web: http://www.emdmillipore.com/CA/en/20160414_183417

www.laboratoryfocus.ca

Laboratory Focus November/December 2016

NOVEMBER 2016 November 22

Calendar

Email: convention@bio.org Web: http://convention.bio.org/2017/

JUNE 2017 June 19-23

JULY 2017

Undertaking Innovation: Canadian Funding ProSpectr’Atom 2017 grams Venue: Quebec City, Canada Venue: Toronto, ON Web: http://www.csass.org/ Web: www.eventbrite.com/e/undertaking-innovaJune 19-22 RC_lab_new:Layout 1 12/19/2012 10:01 AM Page 1 tion-canadian-funding-programs-for-startups-tickets-26636093272 Bio International Conference Venue: San Diego, CA

July 23-26 BIO World Congress on Industrial Biotechnology Venue: Montreal, QC Web: www.bio.org/events/bio-world-congress

November 28

LSO Annual Policy Forum Venue: Toronto, ON Web: www.lifesciencesontario.ca/events/eventsCalendar/PolicyForum2016.php

DECEMBER 2016 SRC103

December 1 Genesis 2016 Venue: London, UK Web: www.genesisconference.com

December 7 Canadian Biomanufacturing Consultation Workshop Venue: Montreal, QC Tel: 514-439-5538 or 1-844-285-2342 Email: info@biomanufacturing.ca Web: www.biomanufacturing.ca

FEBRUARY 2017

I wish to receive/continue to receive a complimentary subscription to

Yes

LABORATORY FOCUS

Format Preference:

Print option

Digital option

Signature:________________________________Date: ____________________________ Name: ____________________________Job Title:_______________________________ Company: _________________________Dept: _________________________________ Business Address : _______________________________________________________

February 4-8

City: _________________________Prov: ________Postal Code: ___________________

SLAS 2017 International Conference & Exhibition Venue: Washington, DC Tel: 1-877-990-7527 or 1-630-256-7527 Fax: +1.630.741.SLAS (7527) E-mail: slas@slas.org Web: www.slas2017.org

Telephone: ___________________________Fax: ________________________________

February 13-14 Bio CEO & Investor Conference Venue: New York, NY Web: www.bio.org/events/bio-ceo-investor-conference/

MARCH 2017 March 5 Pittcon 2017 Venue: Chicago, IL Tel: 1-800-825-3221 Email: info@pittcon.org Twitter: https://twitter.com/Pittcon Web: www.pittcon.org

March 20-22 BIO Europe Spring Venue: Barcelona, Spain Web: https://ebdgroup.knect365.com/bioeuropespring/

MAY 2017 May 26-28 Labcon 2017 Venue: Banff, AB Web: https://labcon.csmls.org/

E-mail: ___________________________________________________________________ On occasion, LABORATORY FOCUS will send third-party information on products & services related to the lab and life science industries. These may be cancelled at any time. Please check here if you do NOT wish to receive these.

JOB TITLE 70 Laboratory Dir. / Mgr. 71 Laboratory Purchaser 72 Laboratory Technician 73 Research Scientist 99 Other:____________________ ____________________________

C75 10 11 12 13 14 15

C87 A B C D

C90

COMPANYs PRIMARY BUSINESS ACTIVITY 50 Industrial Laboratories 51 Academic Laboratories (except medical) 52 Medical Laboratories and Pharmacies within Hospitals and Universities (clinical and research)

55 Government Laboratories 54 Pharmaceutical Companies including Pharmaceutical Wholesalers 57 Private (Independent) Lab 99 Other:____________________ ____________________________

Primary Work Field Aerospace Automotive Biological Sciences Chemicals Electrical/Electronics Energy

16 Environmental Science 17 Food/Beverages/ Agriculture 18 Forensics 19 Genetic Technology 20 Material Science

27 Plastics/Rubber 28 Supplier – Instruments/ Consumables 29 Manufacturing 30 Construction 99 Other: ____________

21 Clinical Sciences 22 Metallurgy 23 Paints/Coatings 24 Paper/Pulp 25 Petroleum 26 Pharmaceuticals

Products Used in your Laboratory Analysis Instruments Basic Lab Equipment Chemicals/Biochemicals Chromatography – Gas

E F G H

Chromatography – Liquid Filtration, Water Purification LIMS Liquid Handling & Sample Prep

I J K L

Microscopes, Optics, Cameras Safety & Hygiene Spectroscopy Testing Systems/Equipment

M Vacuum Equipment

To subscribe to our e-mail newsletters, please check

1 Biotechnology Focus eBulletin 2 Laboratory Focus eBulletin

3 BioPharma 4 BioMedical 5 Health Care

6 Agri-Food 7 Clean Tech 8 Industry Inte

For a quick response please fax: 905-727-4428 or e-mail: circulation@promotive.net

Laboratory Focus November/December 2016 www.laboratoryfocus.ca

app review

Doctor Pocket™

http://www.doctorpocket.ca

Ontario hands out its 2016 Polanyi prizes The province of Ontario recently announced the winners of its 2016 John Charles Polanyi Prizes. The awards recognize up and coming researchers in the fields of physics, chemistry, physiology or medicine, literature and economic science. This year’s winners are: Dr. Stephen Newman, Dr. César Sosa-Padilla, Dr. Daniel Wright, Dr. John Antoniadis, and Dr. Kyle Biggar. The awards are given every year to five researchers who are in the early stages of their careers and pursuing post-doctoral research at an Ontario university. Recipients are considered the province’s next generation of star innovators. A panel directed by the executive heads of the Council of Ontario Universities selected the prize recipients from a collection of nominees. This year’s scientific winners are working on diverse bodies of research, including curing breast cancer and developing new, ‘green’ chemical reactions and processes. Taking home the chemistry prize was Dr. Stephen Newman, an assistant professor and Tier 2 Canada Research Chair in the Department of Chemistry and Biomolecular Sciences at the University of Ottawa. Dr. Newman’s research uses modern technology to enable environmentally safe preparation of pharmaceuticals, materials and bulk chemicals. This year’s physics winner is Dr. John Antoniadis, a Dunlap Fellow at the Dunlap Institute for Astronomy and Astrophysics at the University of Toronto. Dr. Antoniadis’ research project combines theory with practical observations to better understand the properties and diversity of neutron stars, which orbit other stars. Taking top honours in the Physiology/ Medicine category is Dr. Kyle Biggar, an assistant professor at the Institute of Biochemistry at Carleton University. Dr. Biggar’s research focuses on the resistance of breast cancer to chemotherapy, specifically the oxygen supply in tumors, which plays a significant role in cancer progression, aggressiveness and resistance to chemotherapy. Ontario established the Polanyi Prizes in 1987 to honour the achievement of John Charles Polanyi, a 1986 Nobel Prize Laureate in chemistry. Dr. Polanyi won his Nobel Prize 30 years ago.

Need a doctor on call? Imagine having a rich network of medical specialists at your fingertips. Doctor Pocket™ is an iOS and Android application that enables you to communicate with specialists from the comfort of your home and on-the-go. In Canada, everyone may have health insurance, but that doesn’t mean everyone has equal and easy access to care. Doctor Pocket changes that by connecting you with a specialist of your choice to get the medical attention you need within 24 hours. Book and chat with doctors from Harvard, Yale, Johns Hopkins, and McGill. With Doctor Pocket, users can personalize their account by entering basic profile information that will be seen by specialists in family medicine, pediatrics, radiology, and dermatology. Booking the appointment is a breeze: users select a doctor by specialty and pick a date and time to meet. Patients can share high resolution images, videos and voice notes directly from their phone with a simple click.

ChemWizard

From Google https://itunes.apple.com/us/app/chemwizard-balancer-masses/ id1000353235?mt= Developed by students, for students, ChemWizard was developed to assists chemistry students and graduates alike with the tedious task of balancing and calculating values of equations and their molecules. Whether you’re a student looking to speed up your work, a professional trying to be more productive, or just somebody who really struggles with stoichiometry problems, ChemWizard can help. Not only does the app balance chemical equations, it will also calculate the atomic and molar masses of the individual elements and molecules, respectively, within those chemical equations. The app also includes calculators to find the limiting and excess reagents of a chemical equation, as well as to convert grams to moles (and vice versa). Best of all, Chemwizard is supported by all devices including phones and tablets. It’s also available from both the Apple App store and the Google Play store.

SAVE TIME Click here first for the latest information in the laboratory market. Don’t get left behind.

www.laboratoryfocus.ca

@LabFocus

New: 4 liter capacity!

More Capacity Centrifuge 5920 R The new Centrifuge 5920 R delivers extraordinary high capacity in a very compact and ergonomic product design. Its state-of-the-art refrigeration system provides excellent cooling performance and keeps your temperature sensitive samples safe.

> Max. capacity: 4 x 1000 mL or 52 x 50 mL conical > Dual use buckets for tubes and plates > Benchtop centrifuge with floor-standing capabilities > Designed for even lower noise levels

www.eppendorf.com/centrifugation • 800-263-8715 Watch video!

C012.A1.0116.A.CA-LFO.indd 1

Full page tabloid ad Laboratory Focus – LFO

4/18/16 10:54 AM

Laboratory Focus November/December 2016

Articles inside

New Products

App Reviews

Appointments