Laboratory Focus June/July 2016

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June/July 2016 Volume 20, Number 2

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R&D News.......................... 1 Appointments..................... 5 Pharma Notes..................... 7 New Products................... 15 App Reviews...................... 18 Calendar........................... 19

Johnson & Johnson Innovation opens JLABS @ Toronto

TORONTO, ON – Johnson & Johnson Innovation has officially opened its newest JLABS life sciences incubator in Toronto, ON. The new 40,000-square-foot life sciences incubator called JLABS @ Toronto provides entrepreneurs shared lab space and offices, modular lab suites and access to scientific, industry and capital funding experts as they work to build their early-stage companies.

In all, 22 companies focused on therapeutics, medical devices, and

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consumer health solutions were named as the first residents at the facility. The first group of resident startups also includes the seven winners of the JLABS @ Toronto Quick Fire Challenge, which awards promising early-stage innovation companies with residency at the facility. The new facility can accommodate up to 50 startups Johnson & Johnson said.

“The expansion of JLABS into Canada helps further Johnson & Johnson Innovation’s mission to

discover and advance the best science in the world that has the most potential to help patients and consumers,” said Dr. Paul Stoffels, chief scientific officer and worldwide chairman, pharmaceuticals, Johnson & Johnson. “We are confident that JLABS @ Toronto will support a vibrant ecosystem of startups and entrepreneurs with access to the world-class expertise and technology within our global network.”

JLABS @ Toronto is the sixth JLABS facility to open and the first outside of the United States.

JLABS @ Toronto is located at MaRS Discovery District and is a collaboration between Johnson & Johnson Innovation LLC, the University of Toronto, MaRS Discovery District, Janssen Inc., MaRS Innovation and the Government of Ontario, and includes the following hospital partners: Centre for Addic-

tion and Mental Health, the Hospital for Sick Children, Sinai Health System, St. Michael’s Hospital, Sunnybrook Health Sciences Centre and University Health Network.

The new facility will also feature a device and digital prototype lab that will provide entrepreneurs access to highly specialized tools, as well as skill-building programs to design and develop smart health technologies.

Additionally, resident companies at JLABS @ Toronto also have the opportunity to collaborate with IBM Canada to access its entrepreneurship programs and services, including IBM Watson cognitive business technology and IBM Bluemix cloudbased development platform.

“We are thrilled to be part of the flourishing life sciences community in Toronto and to contribute to Ontario’s vision to help drive the province toward becoming a world leader in innovation for health and wellness,” said Melinda Richter, head of JLABS. “Canada’s startup scene is booming, and we look forward to working with the many enterprising innovators in the region that are working to turn science into tangible, commercial products.”

JLABS @ Toronto joins a network of JLABS sites throughout North America in life science clusters, including San Diego (flagship), San Francisco, South San Francisco, Boston and Houston. These facilities are currently home to more than 120 early-stage companies advancing bio/pharmaceutical, medical device, consumer and digital health programs. The six JLABS facilities have a total capacity for 225 resident companies.

To see this story online visit http://www.laboratoryfocus.ca/ johnson-johnson-innovationopens-jlabs-toronto/

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news

Canada’s top student scientist recognized at Sanofi Biogenius Canada national final OTTAWA, ON – Iveta Demirova from New Westminster Secondary School in New Westminster, BC has been awarded top honours at the

national final of the prestigious Sanofi Biogenius Canada (SBC) competition in Ottawa.

The 16-year-old, grade 11 student, was chosen by the

judges for her research project exploring the development of a novel HIV-1 therapy.

The results of Iveta’s research project, completed

with the support of mentor Dr. Ralph Pantophlet of Simon Fraser University, could offer Continued on page 3

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June/July 2016

news Continued from page 2 numerous advantages to those living with HIV, which remains one of the world’s leading infectious diseases. Although current treatment options have managed to successfully target and suppress the virus among patients, many individuals become resistant to treatment, and it is among this population that Iveta’s project could play a significant role.

“I was very pleased with the results of my research and I am hoping that my findings will have an impact within the field of HIV research, and, more importantly, in the lives of patients living with this disease,” she said. “I am truly honoured to have won the Sanofi Biogenius Canada competition, and thrilled to have the opportunity to represent the country at the 2016 International BioGENEius Challenge in San Francisco in June.”

One of the country’s most prestigious student competitions, Sanofi Biogenius Canada pairs exceptional young scientists at the high school level with academic mentors to pursue real-world research projects. These enriching partnerships have resulted in

Iveta Demirova

many promising breakthroughs across various scientific fields.

Hosted at the National Research Council of Canada (NRC), the competition’s national final featured the winners of all nine Sanofi Biogenius

Canada regional competitions across the country. The nine finalists presented their research projects to a judging panel of esteemed members of the scientific community, including Dr. Robert Tsushima, associate dean, Research and Partnerships, Faculty of Science, York University; Dr. Thomas Merritt, Canada Research chair in Genomics and Bioinformatics, Laurentian University; Dr. Lakshmi Krishnan, program Leader, NRC and Michael McCluskie, senior research officer, NRC. Iveta receives a cash prize of $5,000, a portion of which will go to New Westminster Secondary School, and she will now progress to the 2016 International BioGENEius Challenge in San Francisco in June, where she will submit her work to a panel of preeminent international scientists.

Runners-up in this year’s national SBC competition were awarded cash prizes ranging in value from $1,000 to $4,000.

Second prize went to Melody Song, a student from Evan Hardy Collegiate in Saskatoon, SK. Melody earned high praise for her research project which focused on preventing disease in Faba

Bean crops, and was completed with the support of mentors Dr. Kirstin E. Bett and Dr. Hamid Khasaei of the University of Saskatchewan.

Third prize was awarded to Denis Drewnik, a grade 12 student from Sisler High School in Winnipeg, MB, for a project that examined how to protect the canola crop from the devastating Blackleg disease, which can reduce yield by up to 20 per cent and have a devastating economic impact. Denis’ project was supported by mentor Dr. Mark Belmonte of the University of Manitoba.

Dina Shehata from Holy Heart of Mary High School in St. John’s, Newfoundland earned the competition’s Commercialization prize. The Commercialization prize recognizes the project with the most commercial potential and viability. Dina’s research focused on developing a low cost gel model for ultrasound training.

To see this story online visit http://www.laboratoryfocus.ca/ canadas-top-student-scientist-recognized-at-sanofi-biogenius-canadanational-final-2/

Scientists map the spread of deadliest ovarian cancer Dr. Sohrab Shah

VANCOUVER, BC – BC Cancer Agency researchers are providing critical insight into the invasive spread of the most malignant form of ovarian cancer in a landmark study, published in Nature Genetics. This is a first in mapping two distinct patterns of ovarian cancer cell migration in high grade serous ovarian cancer.

The discovery, led by Dr. Sohrab Shah, senior scientist at the BC Cancer Agency, associate professor at the University of British Columbia, and Canada Research Chair in Computa-

tional Cancer Genomics, was made possible through genomic sequencing techniques and novel software developed by his bioinformatics team at the BC Cancer Agency. The study was simultaneously published in Nature Methods. The scientists have answered key unknowns about how deadly ovarian cancers spread, and the composition of the cancer cell groups that have taken up residence within the patient’s abdomen.

The study reveals that many cancer cell types make up a patient’s tumour.

This could explain why some cells are susceptible to treatment when others are resistant, leading to relapse. Also, cell type migration patterns from ovary to other abdominal sites identified that specific ovary sites contained many more cell types relative to others. These regions could pinpoint ‘gateways’ of cell migration to other abdominal sites.

Unlike most cancers that spread through the blood stream or lymph system, this study shows that high grade serous ovarian cancer cells have a unique opportunity to spread

prolifically throughout the abdomen.

In mapping the cell migration, Dr. Shah’s team shows how cells are able to settle and thrive in specific regions of the body causing widespread, lifethreatening disease.

The migration maps were primarily determined in pre-treatment samples with one exception. Shah’s team studied one patient with multiple relapse specimens. Notably, only a subset of cancer cells present at diagnosis led to treatment resistance.

The next steps are to use the techniques developed for this study to define cell migration maps from additional patients over time with a specific focus on determining which cells are resistant to treatment. This will allow researchers to build predictive tools to better inform future care.

Moreover, this new understanding of how high grade serous ovarian cancer cells migrate within the patient’s body provides insight that could inform future treatment selection. These results indicate that some cancer cells may have had pre-existing properties of resistance prior to the patient taking any treatment and could indicate that a patient requires a much more aggressive, multi-treatment approach from the start to prevent relapse.

Dr. Shah’s work is supported by the BC Cancer Foundation.

To see this story online visit http://www.laboratoryfocus.ca/ scientists-map-the-spread-of-deadliest-ovarian-cancer/

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news

Researchers discover blood marker that better defines response to drug used for metastatic colorectal cancer

Dr. Geoffrey Liu TORONTO, ON – Cancer researchers have identified a marker that shows up in a blood test that determines which patients with colorectal cancer that has spread would benefit from receiving the drug cetuximab.

The research, published online in Clinical Cancer Research, a journal of the American Association for Cancer Research, solves one of the mysteries of why receiving cetuximab was futile for up to half of incurable colorectal cancer patients not responding to treatment. The research was conducted by principal investigator Dr. Geoffrey Liu, a clinician-scientist at Princess Margaret Cancer Centre, University Health Network, in collaboration with the Canadian Cancer Trials Group and the Australasian GastroIntestinal Trials Group.

“Our research discovered that the blood marker FCGR2A identifies a new group of patients that will benefit from taking cetuximab,” says Dr. Liu. “With this finding, we believe we are now on the way to move it into the clinical setting to provide patients targeted, effective treatment.”

The new research builds on an international clinical trial of about 10 years ago led by Canadians, in which the Princess Margaret participated.

“In a group of metastatic colorectal cancer patients who were running out of treatment options, the previous clinical trial determined that cetuximab was most effective in a certain group of patients with tumours carrying a RAS mutation. But it certainly didn’t work for everyone and so the race was on to find out how to better identify which patients would benefit from this drug,” says Dr. Liu. “Our finding, which resulted from analyzing archived tumour and normal tissue samples from some of the 572 patients enrolled in that trial, further refines this quest and defines another subset of patients who will respond to the drug,” he said. “We need to find other ways to personalize cancer medicine for people with colorectal disease, keeping in mind that cetuximab is an expensive drug and can have side effects. So instead of looking at aspects in the tumour, which is where RAS mutations show up, we looked at certain things in the blood and normal tissues that we

could measure for heritable genetic variations.”

Dr. Liu, a medical oncologist who specializes in lung cancer, describes the pathway to the discovery as an ideal example of what can happen when you follow the drug and wherever the research may lead.

“When we followed the drug, first in lung cancer and then in other cancers, it led us to colorectal cancer where the drug was also being used, and directly onto this new finding.”

Dr. Liu’s research was funded by the Ontario Institute of Cancer Research, the Alan B. Brown Chair in Molecular Genomics, the Cancer Care Ontario Chair in Experimental Therapeutics and Population Studies, the Canadian Cancer Society, and The Princess Margaret Cancer Foundation.

To see this story online visit http://www.laboratoryfocus.ca/ researchers-discover-blood-markerthat-better-defines-respond-todrug-used-for-metastatic-colorectalcancer/

McMaster scientists uncover new way to grow rare life-saving blood stem cells HAMILTON, ON – Researchers at McMaster University’s Stem Cell and Cancer Research Institute have taken a significant step forward in understanding the stem cells of the human blood system after discovering how a key protein allows for better control and regeneration of these cells.

This discovery, published in the scientific journal Nature, illustrates how a protein called Musashi-2 regulates the function and development of important blood stem cells. This knowledge provides new strategies that can be used to control the growth of these cells — cells that can be used as therapeutics for a range of life-threatening diseases but are, in general, in very short supply.

The senior author of the study is Kristin Hope, principal investigator at the Stem Cell and Cancer Research Institute and assistant professor with McMaster University’s Department of Biochemistry and Biomedical Sciences. The research also involved collaborators from the University of California San Diego, University of Toronto and the University of Montreal.

Hope says the discovery could

be impactful for the tens of thousands of patients suffering from a range of blood-based disorders including leukemia, lymphoma, aplastic anemia, sickle cell disease and more.

“We’ve really shone a light on the way these stem cells work,” she says.

“We now understand how they operate at a completely new level, and that provides us with a serious advantage in determining how to maximize these stem cells in therapeutics. With this newfound ability to control the regeneration of these cells, more people will be able to get the treatment they need.”

The research team specifically looked at stem cells from umbilical cord blood, a proven but under-utilized source of stem cells for the treatment of adult blood cancers. These stem cells have the potential to become an important therapeutic for the thousands of people suffering from blood cancers who are awaiting life-saving transplants.

Cells from umbilical cord blood have unique properties that make them easier to use for transplantation, including accessibility and adaptability.

As a result, they allow for safer and more effective transplants.

The problem, Hope points out, is

Kristin Hope

that there are very few stem cells available in individual cord blood samples — only about five per cent of all samples actually contain enough cells for a transplant. The team’s research into the importance of Musashi-2 and its role in expanding the number of stem cells in a given cord blood sample could help ease the current stem cell shortages.

Gene Yeo, associate professor at the University of California San Di-

ego, co-corresponding author of the study, adds, “Most stem cell studies focus on proteins that bind DNA to control gene output.

The prominent role we found for Musashi-2, a protein that instead binds to RNA, also underscores an urgency to study this second layer of gene regulation in stem cells.”

Hope says: “Providing enhanced numbers of stem cells for transplantation could alleviate some of the current post-transplantation complications and allow for faster recoveries, in turn reducing overall health care costs and wait times for newly diagnosed patients seeking treatment.”

“By expanding the stem cells as we have done, many more donated samples could now be used for transplants.”

The study was supported by the Ontario Institute for Cancer Research, the Canadian Institutes of Health Research, Canadian Blood Services, Health Canada, the National Institutes of Health and the California Institute for Regenerative Medicine.

To see this story online visit http://www.laboratoryfocus.ca/ mcmaster-scientists-uncovernew-way-to-grow-rare-life-savingblood-stem-cells/

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Laboratory Focus June/July 2016

news Backed by CIHR, Ontario team to study metabolic syndromes TORONTO, ON – Dr. Philip Awadalla, a senior investigator at the Ontario Institute for Cancer Research (OICR) and principal investigator for the Ontario Health Study has been awarded $2 million by the Canadian Institutes of Health Research (CIHR).

The award will fund the study of the role of both genes and the environment on the development of metabolic syndrome, a cluster of medical conditions that are common in aging adults, including obesity, hypertension, high cholesterol, high blood sugar and insulin resistance. These conditions are considered to be both risk factors and causal factors in the development of cancer and chronic diseases like heart disease, stroke and diabetes. A better understanding of how to prevent and treat the conditions of metabolic syndrome could also help in the design of new strategies to prevent these diseases before they develop.

Awadalla and his team will use data and samples from the Ontario Health Study, which is part of the larger Canadian Partnership for Tomorrow Project, to conduct their research.

“There are currently no research projects at this scale looking at the genetic and environmental risk factors of metabolic syndrome or related aging-associated traits, which affects close to one quarter of Canada’s adult population,” said Dr. Awadalla.

“This funding will allow us to tap into the vast data holdings of the Ontario Health Study to investigate the interplay among varying environmental exposures, aging, and epigenetic factors across Ontario and Canada. Ultimately we hope to find new ways to better manage these conditions and other linked diseases such as cancer.”

The team has partnered with Illumina to assist with genome sequencing, as well as Environment Canada and the Canadian Partnership Against Cancer.

To see this story online visit http://www.laboratoryfocus.ca/ backed-by-cihr-ontario-team-tostudy-metabolic-syndromes/

Chance finding from University of Guelph study could transform plant production The wild plant is on the left and the transgenic plant on the right.

Photo credit: University of Guelph

GUELPH, ON – An almost entirely accidental discovery by University of Guelph researchers could transform food and biofuel production and increase carbon capture on farmland.

By tweaking a plant’s genetic profile,

the researchers found a way to double the plant’s growth and increased seed production by more than 400 per cent.

The findings were published in the March 2016 issue of Plant Biotechnology Journal.

The team studied Arabidopsis, a small flowering plant often used in lab studies because of its ease of use and its similarity to some common farm crops. They found that inserting a particular corn enzyme caused the plant’s growth rate to skyrocket.

“Even if the effects in a field-grown crop were less, such as only a tenth of what we’ve seen in the lab, that would still represent an increase in yield of 40 to 50 per cent, compared with the average one to two per cent a year that most breeding programs deliver,” said Prof. Michael Emes, Department of Molecular and Cellular Biology (MCB).

He said the team’s finding could boost yields of important oilseed crops such as canola and soybean, as well as crops such as camelina, increasingly grown for biofuels. Larger plants would capture more atmospheric carbon dioxide without increasing the amount of farmland, said Emes. “Farmers and consumers would benefit significantly in terms of food production, green

Canada and Japan usher in a new era of partnership in physics research

(L to R) Dr. Jonathan Bagger, Director of TRIUMF, Canada’s Minister of Science Kirsty Duncan and Dr. Masanori Yamauchi, Director General of KEK Photo Credit: TRIUMF (TSUKUBA, JAPAN/VANCOUVER, BC) – On May 15, Canada’s Minister of Science Kirsty Duncan welcomed a new era of world-class scientific partnership between Canada and Japan as she unveiled the new TRIUMF branch office located at Japan’s KEK. Minister Duncan was joined by dignitaries from both laboratories to perform the ribbon cutting, celebrating the research collaboration between these two hubs for subatomic physics research.

The new branch office, which is also shared with CERN, follows the recent signing of a new partnership

agreement this past December by Dr. Jonathan Bagger, director of TRIUMF – Canada’s national laboratory for particle and nuclear physics and acceleratorbased science – and Dr. Masanori Yamauchi, director general of KEK – a high energy accelerator research organization in Japan. The agreement enhances research collaborations between the two labs to answer questions on areas ranging from the breadth and composition of the universe to topics closer to home, such as the properties of advanced materials.

“As world leaders in subatomic physics, TRIUMF and KEK have forged

energy and the environment. The ramifications are enormous.”

The finding came almost by chance.

Studying the enzyme’s effect on starch, the researchers noticed that their genetically engineered plants looked different and much larger in photos taken by lead author Fushan Liu, a former post-doctoral MCB researcher.

“That’s when we realized that we were looking at something potentially much more important,” said Ian Tetlow, an MCB professor and study co-author.

Although genetic engineering led to more flowers and pods containing seeds, it left the seed composition unchanged.

“The seeds are where we would get the oil from, and consistent composition is important so that the function and use of the oil isn’t changed,” said Tetlow.

The researchers plan to test canola and other crops. Field tests and analysis with industry and government will likely take several years.

To see this story online visit http://www.laboratoryfocus.ca/ chance-finding-could-transformplant-production-u-of-guelph-study/ an extraordinary collaboration that continues to unlock new opportunities to advance this important field,” said Duncan.

“For decades, TRIUMF and KEK have been recognized internationally in the areas of subatomic physics, accelerator science and materials science,” said Yamauchi. “Through our growing partnership, we will continue to be global leaders in advancing these areas of research, as well acting as pillars of scientific co-operation.”

“The opening of this new branch office represents not just a strengthening of the partnership between TRIUMF and KEK, but also the importance of collaboration on the global scale,” adds Dr. Jonathan Bagger, TRIUMF director.

TRIUMF and KEK have numerous shared projects in the areas of subatomic physics, accelerator science, and materials science. Current efforts include the T2K and Belle II experiments in Japan, the Large Hadron Collider at CERN, and the proposed International Linear Collider The hope of this new office and indeed the new partnership agreement is to advance scientific discovery through enhanced bilateral collaboration.

To see this story online visit http://www.laboratoryfocus.ca/canada-and-japan-usher-in-a-new-era-ofpartnership-in-physics-research/

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June/July 2016 Laboratory Focus www.laboratoryfocus.ca

Appointments

Rna Diagnostics Inc. has named Dr. Jeremy Bridge-Cook as its new president and CEO. Dr. Bridge-

Vitae Pharmaceuticals and Idenix Pharmaceuticals.

John Sebben has joined Purdue as vice president, manufacturing operations. Previously he worked at a number of pharmaceutical companies including: GlaxoSmithKline, Apotex, Teva Canada, Pharmascience, Biovail and most recently, Patheon, where he held the role of vice president, North American supply chain. Sebben holds an MBA from York University and BSc from University of Toronto.

Dr. Jeremy Bridge-Cook

Cook is a highly accomplished In Vitro Diagnostics (IVD) executive with experience in developing and launching innovative IVD products onto global markets. He has held progressive senior business development, research and development and general management roles at Tm Bioscience (TMB) and Luminex Corporation, which acquired TMB in 2007. He was also a corporate officer and senior vice president of R&D at Luminex. Dr. Bridge-Cook obtained his B.Sc., Biology from McMaster University and his Ph.D., Immunology from University of Toronto.

Aurinia Pharmaceuticals Inc. has appointed Charles Rowland as its new Chief Executive Officer replacing Stephen Zaruby who has announced his resignation from the position and and from the company’s board of directors. Rowland’s last role was vice president and chief financial officer of ViroPharma Incorporated prior to its acquisition by Shire in January 2014. Prior to joining ViroPharma in 2008, he held a number of leadership positions at several biotechnology and pharmaceutical companies, most recently as interim co-CEO, executive vice president and chief financial officer for Endo Pharmaceuticals Inc. He also held positions of increasing responsibility at Biovail Corporation, Breakaway Technologies Inc., Pharmacia Corporation, Novartis AG and Bristol-Myers Squibb. He has board experience with companies such as BluePrint Medicines,

Pascal Spothelfer has been named by Genome British Columbia (Genome BC) as its new president & CEO beginning Monday, June 6. He joins Genome BC with a wealth of international experience including time as a management consultant with the Boston Consulting Group and as an executive of Jenoptik AG in Germany. After four years as president and CEO of NovAtel Inc. in Calgary, Spothelfer moved to Vancouver where his local background includes time as president and CEO of Spectrum Signal Processing, over three years at the helm of the BC Technology Industry Association (BCTIA) and vice president communications and community partnerships at the University of British Columbia.

Spothelfer holds a law degree and a PhD in Law from the University of Basel in Switzerland and a MBA from INSEAD in Fontainebleau, France. The appointment follows the retirement announcement of Dr. Alan Winter late last year. Winter will continue in his role of president & CEO until June 6th and help with the transition until mid-July 2016.

Lutz Goedde, a leading expert in strategies to improve agricultural productivity around the world, will join the board of directors of the Global Institute for Food Security (GIFS) at the University of Saskatchewan (U of S). Goedde is a partner in the Denver office of McKinsey & Company, where he is a leader in the firm’s agriculture/ food and social sector practices.

He has worked with agriculture and food companies, governments, and NGOs in North America, India, Europe, and Africa to improve agricultural productivity and efficiency, increase food availability and supply chains, and reduce poverty and

hunger. He was part of the leadership team that built the global and agricultural development program at the Bill & Melinda Gates Foundation. During his tenure, the Gates Foundation committed over $2 billion to development efforts in Africa and Asia to improve smallholder agriculture.

Life Sciences Ontario (LSO) has welcomed Rebecca Yu, head of JLABS @ Toronto at Johnson &

Rebecca Yu Johnson Innovation, to its board of directors. A leading life sciences and health technology innovator, Ms. Yu was a key player behind bringing the successful JLABS incubator to Toronto – the first outside the U.S.. Ms. Yu will head the 40,000-square foot Toronto facility, a collaboration between Johnson & Johnson Innovation, The University of Toronto, MaRS Discovery District, Janssen Inc., and MaRS Innovation, with support from the Government of Ontario. Prior to her role with JLABS, she was director, Strategic Health Technology Assessment (HTA), Government Affairs and Market Access, Janssen Inc., one of the Janssen Pharmaceutical companies of Johnson & Johnson, where she led HTA policy strategy on behalf of Janssen, including work with HTA agencies/ bodies on incorporating global best practices. Prior to joining Janssen in 2012, Yu enjoyed a 15-year career in the pharmaceutical industry, taking on progressively senior roles in the areas of health policy, medical affairs, and government affairs with Solvay Pharma, Procter & Gamble, and Pfizer. She is a graduate of the University of

Toronto’s Faculty of Pharmacy, and a licensed and practicing pharmacist in Ontario.

Zymeworks has named Dr. Diana Hausman as its new chief medical officer. In this role, Dr. Hausman is responsible for overseeing clinical development and clinical strategy for the company. A board certified medical oncologist, Dr. Hausman brings more than 15 years of clinical drug development experience to the role. Most recently, she was chief medical officer at Oncothyreon where she oversaw the clinical program for their lead Phase 2 targeted anti-HER2 cancer therapy. She has also held positions at Zymogenetics, Berlex, and Immunex working across multiple indications, including oncology, hematology, hepatitis C, and autoimmune disease. Dr. Hausman received her internal medicine training and specialty training in hematology and medical oncology at the University of Washington.

She holds an M.D. degree from the University of Pennsylvania and an A.B. in biology from Princeton University.

Oncolytics Biotech® Inc. has announced the formation of a Science and Technology Committee made up of directors Drs. William Rice and Bernd Seizinger. Dr. William Rice has held the position of chairman, president and CEO of Aptose Biosciences Inc. since 2013.

Preceding that, he served as chairman, president and CEO of Cylene Pharmaceutics Inc. He has also served as senior scientist and head of the Drug Mechanism Laboratory at the National Cancer InstituteFrederick Cancer Research and Development Center, and as a faculty member in the Division of Pediatric Hematology and Oncology at Emory School of Medicine. Dr. Seizinger has held the position of chairman/ executive chairman of Opsona Therapeutics Ltd. since 2009.

From 1998 to 2009, he served as president and CEO of GPC Biotech.

He also served as vice president of Oncology Drug Discovery and, in parallel, vice president of Corporate and Academic Alliances at Bristol-Myers Squibb. He has also held professorships and senior staff appointments at Harvard Medical School, Princeton University and Massachusetts General Hospital.

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Laboratory Focus June/July 2016

Pharma Notes

Health Canada has given ProMetic Life Sciences (Laval, QC) CTA clearance to commence its Phase 2 clinical trial for its orally active anti-fibrotic lead drug candidate, PBI-4050, in patients suffering from cystic fibrosis (CF). The objectives of the 24 week randomized, double-blind, and placebo-controlled Phase 2 study includes the evaluation of the effects of PBI-4050 on pancreatic and lung function in 90 CF patients. To date there have been no drug-related serious adverse events and PBI-4050 has been very well tolerated by patients.

Eupraxia Pharmaceuticals (Victoria, BC) has initiated STEP UP – a Phase 1 study for its lead investigational product, EP-104IAR, for the treatment of pain associated with osteoarthritis (OA) of the knee. EP-104IAR is an extended release formulation of the commonly used corticosteroid fluticasone propionate that is delivered by intra-articular injection into the knee. Eupraxia’s patented technology coats the drug particle with a biocompatible polymer that has been shown to extend drug release into the knee joint for our target of six months and beyond, while limiting systemic exposure. STEP UP is a multi-centre, double-blind, placebocontrolled clinical trial that will enroll 32 patients with moderate to severe knee OA between the ages of 40 to 80 years. Patients will receive a single 15 mg injection of EP104IAR (or placebo) in one affected knee and be followed for up to 42 weeks or until their knee returns to its prior level of pain. The main purpose of the study is to profile the safety and pharmacokinetics of EP104IAR, and may also provide some preliminary insight into the duration and degree of pain relief. The trial, which has a Health Canada CTA approval, will be conducted across two sites in London, ON by Dr. Getgood, associate scientist, and Dr. Rob Petrella, scientist, of the Lawson Health Research Institute.

Results are anticipated in 2017.

Theralase Technologies Inc. (Toronto, ON) has signed a clinical research agreement with the Princess Margaret Cancer Centre, University Health Network (UHN) to conduct a Phase 1b clinical study for the indication of non-muscle invasive bladder cancer

(NMIBC). Theralase is a biotech company looking to commercialize medical devices to eliminate pain and a developer of photo dynamic compounds to destroy cancer. The clinical study entitled, “A Phase Ib Trial of Intravesical Photodynamic Therapy in Patients with NonMuscle Invasive Bladder Cancer at High Risk of Progression Who Are Refractory to Therapy with Bacillus Calmette-Guerin (“BCG”) and Who Are Medically Unfit for or Refuse a Cystectomy”, will be conducted by the department of urology at UHN, under the guidance of clinical principal investigator Dr. Girish Kulkarni.

Cardiome Pharma Corp.

(Vancouver, BC) through its affiliate Cardiome Group has signed an exclusive license agrreement with an affiliate of Allergan plc that will result in Cardiome commercializing XYDALBA™ (Dalbavancin) in France, the U.K., Germany, Belgium, Nordic nations, certain other European nations (not already partnered), various Middle Eastern nations and Canada. Cardiome will provide Allergan with a staggered upfront payment totaling US$13 million and additional milestone payments and royalties based upon

commercial achievements and sales of XYDALBA™. Additional terms of the agreement were not disclosed.

Transition Therapeutics Inc. (Toronto, ON) has dosed the first patient in the Phase 2 study of its selective androgen receptor modulator (SARM) drug candidate TT701.

The Phase 2 study will evaluate the efficacy and safety of TT701 in improving the symptoms of androgen deficiency (sexual symptoms, fatigue/low vitality, and physical dysfunction) in men with prostate cancer who have undergone radical prostatectomy for organ-localized prostate cancer. Brigham and Women’s Hospital (BWH) is conducting the investigator-led Phase 2 clinical study which is expected to enroll up to 125 subjects at selected specialized clinical sites including BWH. The principal investigator for the Phase 2 study is Dr. Shalender Bhasin, director of the Research Program in Men’s Health: Aging and Metabolism at BWH and an internationally recognized endocrinologist with expertise in testosterone biology and men’s aging.

Zymeworks Inc. (Vancouver, BC) has struck a second partnership deal with pharmaceutical heavy-

7

weight GlaxoSmithKline (GSK), for the research, development and commercialization of bispecific antibodies. Under the agreement, GSK will have access to Zymeworks platform technology, Azymetric™.

The platform will be employed to develop new multiple bi-specific drugs across different disease areas. Zymeworks will receive upfront and preclinical payments of up to USD $36 million and is eligible to receive up to USD $152 million in development and clinical milestone payments, along with commercial sales milestone payments of up to USD$720 million, and tiered royalties on potential sales. As previously announced in December 2015, Zymeworks and GSK entered into a collaboration and license agreement to further develop Zymeworks’ Effector Function Enhancement and Control Technology (EFECT™) platform and to research, develop, and commercialize novel Fc-engineered monoclonal and bi-specific antibody therapeutics that have been optimized for specific therapeutic effects. As part of this second agreement, GSK has also gained the right to combine the Azymetric™ platform with novel engineered Fc domains developed under the previously announced collaboration.

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b y Roberto Cerruti, Gl ob al Mar k eting Manager , Inst rumentat io n Vacuum, Agilent

Converting to

Oil-free Laboratory Vacuum

A

key element of the GC/ MS instrument is the vacuum system. All instruments equipped with a turbomolecular high vacuum pump require a forepump to assist the turbopump in achieving pressure in the analyzer that is sufficiently low to allow the instrument to work. Until recently, GCMS users have not had an affordable alternative to the low-cost oil-sealed rotary vane pump that has been the traditional device for this purpose. Oilsealed pumps have inconvenient maintenance requirements, introduce the risk of spills, and may even cause contamination due to oil vapor migration. Diaphragm pumps, which are oil-free, have been available as alternatives but are significantly more expensive and introduce an additional set of maintenance issues.

COil Sealed Rotary Vane Pump 1 C Figure 1

Sample Spectra: Oil-sealed Rotary Vane Pump vs. IDP-3 Scroll Pump

RVP RVP

Scroll Pump 1

IDP3 Pump IDP3 Pump

Eliminating Oil, Simplifying Maintenance in the Lab Recently, Agilent has re-visited this challenge, as GCMS instrument users have expressed a desire to simplify management of the pumps, extend maintenance cycles, and eliminate oils from the instrument vacuum system, as well as from the lab in general. The emergence of dry scroll pump technology presents a significant opportunity to meet these objectives.

Dry Scroll Pumps provide the vacuum performance required at comparable cost to oil-sealed pumps, while greatly simplifying maintenance and eliminating oil in the system and the lab.

How Dry Scroll Pumps Work Dry Scroll Pumps consist of an orbiting rotor and a stator, with spiral channels machined into both surfaces. As the moving plate orbits, it traps a volume of gas, and progressively compresses the gas into ever smaller volumes as the “involute” of gas is moved towards the center of

The Mass Assignments and relative intensities are consistent between the RVP and the IDP-3 equipped instrument.

the pump. Unlike rotary vane pumps, which use a film of oil to maintain gas tightness, dry scroll pumps use tight tolerances, and sliding gaskets (“tip seals”) to prevent gas from migrating back towards the pump inlet.

Maintenance Comparison The Scroll Pump requires replace-

ment of two tip seals and a single oring after approximately one year of continuous use. This simple maintenance procedure is clean, and can be accomplished by anyone with modest mechanical skills in less than 30 minutes. (See a video of IDP-3 Tip Seal Replacement @ https://www.youtube.com/watch?v=vUyZb1SWAeE)

Diaphragm Pumps require a full re-build on “at least annual” basis, to prevent the possibility of a diaphragm rupturing, venting the foreline to atmosphere and potentially damaging the turbo-pump. Rebuilding a diaphragm pump is vastly more complex than a tip seal replacement, and may require specialized tools

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Laboratory Focus June/July 2016

Feature Figure 2

Recovery Time After ‘Column Flush’: RVP vs 5x IDP-3

and processes. Diaphragm pump customers typically choose to replace the pump with a re-built version from Agilent rather than tackle the maintenance procedure.

Oil-sealed Rotary Vane pumps require that the oil be removed, replaced and disposed of whenever the oil becomes discolored – typically every 6 to 12 months depending on the application. Exposure to customer samples in many cases will degrade the oil quality in as little as 3 to 6 months, so that far more frequent service intervals are required than with the oil-free IDP-3 Scroll Pump or even the diaphragm pump. Cost of disposal of the used oil, often higher cost per liter than the purchase price of the oil, is also a significant factor, as is the intangible cost of exposing personnel to potentially harmful oil and the inevitable mess and clean up following an oil change.

the forepump to rapidly vent to atmosphere. Exposure to a huge spike in foreline pressure can stop the Turbo Pump instantly, potentially causing irreparable damage.

Rotary vane pumps typically fail through oil starvation (often because the oil has leaked from the pump) or from blocked internal oil lubrication passages in the pump (as a result of contamination or oil degradation).

While virtually all rotary vane pump manufacturers include an oil suckback prevention device, these simple mechanisms are not completely foolproof, and there are many documented cases of catastrophic oil migration back into the analyzer chamber. Repair costs vary, but reports exceeding $10K USD are not uncommon, not including the replacement of the Turbo Pump (which cannot be cleaned in the field) or the Ion Detector which must be replaced.

Failure Mode: Predictable vs.

Catastrophic

IDP-3 VPI Positive Shut-off Valve

As the tip seals wear on a scroll pump, the ultimate (minimum or “base”) pressure that the pump can achieve will slowly begin to rise. This rise is typically not observable by the GC-MS operator, and so may be managed easily through scheduled maintenance. If observed, it will be because an instrument with worn tip seals may take longer for the turbopump to ramp up to full speed, indicating the need for tip seal replacement.

The wear out of a diaphragm pump is far less predictable, and typically maintenance is unplanned. In addition, unless a costly external shut-off valve is fitted, the failure mechanism of a diaphragm pump can have much greater consequences: a rupture of the diaphragm material will cause

The Agilent IDP-3 Scroll Pump models specified for field upgrades of Agilent GCMS instruments include a positive shut-off valve at the pump inlet.

Operation of the valve is controlled by the pump to prevent any chance of foreign particles entering the foreline and potentially causing damage to the instrument. When the pump is switched off (or a power outage occurs), the valve will close immediately, maintaining vacuum in the turbo pump foreline and allowing the instrument to shut down safely.

Analytical Performance To fully compare IDP-3 Scroll Pump’s performance against the oil-sealed rotary vane pump, we considered analytical performance, vacuum performance and audible noise (a critical

Figure 3

Foreline Pressure Recovery After ‘Column Flush: RVP vs IDP-3

factor in customer satisfaction). To confirm the analytical performance of the IDP-3 Scroll Pump on the GCMS instruments we looked at the complete manufacturing test criteria for the instrument, as would apply for those equipped with oil-sealed pumps. Background spectra for the two pump options are shown below. In every analytical performance test conducted, the IDP-3 Dry Scroll Pump delivered equal or better results than its oil-sealed counterpart.

And in certain “ultra-clean” applications, customers may see superior performance from their GC-MS systems with the elimination of oil migration from the rotary vane pump.

Vacuum Performance: Recovery The ability of the GCMS’s forepump to evacuate gas after a sample injection is directly related to the instrument’s sample throughput. As a worst-case scenario, the ability of the IDP-3 Scroll Pump to return the instrument to baseline Analyzer and Foreline Pressures following a 30 ml/ min He column flush was examined.

The time for the Analyzer and Foreline Pressures to return to base (flow = zero) level was consistent between the performance of the Rotary Vane Pump and the IDP-3 pump.

Audible Noise Audible noise produced by the forepump in the GC-MS is typically the largest contributor to the overall noise of the instrument and directly impacts the operator’s work environment. When the average sound level of five IDP-3 Scroll Pumps was compared against a selection of oilsealed rotary vane pumps, the results showed the IDP-3 was consistently 2 dBA lower in noise level at

three of four relative positions, with inconclusive measurements at the fourth. The logarithmic nature of the decibel scale suggests that a 2 dB reduction in audible noise at these levels is readily appreciated by the instrument operator.

The oil-free IDP-3 Scroll Pump provides a turn-key, affordable upgrade to Oil-Free technology for most Agilent GCMS customers. Although there will be some variation by application, Agilent VPD expects GCMS customers to exceed the 1 year recommended replacement interval for IDP-3 Tip Seals. By performing this simple, modestly priced upgrade to their Turbo Pumped GC-MS instruments, customers can access environment-friendly, low ownership cost, dry-pump technology and eliminate messy oil changes and the need for used oil disposal.

Links: •

•

Dry Pump Option for Agilent 5977B lp: http://www. agilent.com/en-us/products/ vacuum-technologies/primarymedium-vacuum-pumps/dryscroll-pumps/idp-3/idp-3-drypump-option-for-5977b-gc-msd IDP3 Speed Curve: http://www. agilent.com/en-us/products/vacuum-technologies/primary-medium-vacuum-pumps/dry-scrollpumps/idp-3/pumpingspeed

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b y And rea C a rus o a nd Ma ss imo S a ntoro, Thermo Fisher Scient ific, Mil an, Italy

Rapid and Reliable Detection of Dissolved Gases in Water Figure 1

To describe a rapid and reliable gas chromatography method for identifying and quantifying methane, ethane, ethylene, and propane in waters affected by hydraulic fracturing operations.

Introduction Hydraulic fracturing, or fracking, is a well stimulation technique in which water is typically mixed with sand and chemicals and injected at high pressure into a wellbore to create small (generally less than 1 mm) fractures to maximize fluid removal and well productivity. This technique enables the removal of large amounts of formerly inaccessible hydrocarbons.

However, the gases liberated using this method may contaminate water reservoirs and have become a controversial environmental and health matter. Monitoring the gas content of affected waters is one way to assess the environmental impact of fracking on an area.

The U.S. Congress has instructed the U.S. Environmental Protection Agency not to regulate hydraulic fracturing, thus some individual states are developing their own regulations. Recently, several states finalized analysis requirements for baseline environmental monitoring of waters prior to hydraulic fracturing. This baseline monitoring aims to identify local water contaminants so that changes can be assessed post hydraulic fracturing to conclusively demonstrate that hydraulic fracturing alone (and not another process, such as mining) was responsible for detrimental impacts on the water.

Different approaches are available for measuring the gas content in water. The most common method is that outlined by the industry standard SOP RSK-175,1 which tests for dissolved gases via static headspace gas chromatography and then calculates results according to Henry’s Law, converting the concentration in the headspace into the partial pressure of the gas and using it to calculate the aqueous gas concentration.

Another possible approach, adopted in this application note, is to saturate water with a gas standard or a mix of gases and use that water to build a calibration curve via sequential dilution. This approach is currently being validated by the American Society for Test Methods (ASTM).

Sample and Standards Preparation Water samples are collected directly

into a 20 mL headspace vial without leaving any headspace. Cap vials immediately and remove 10 mL of water immediately before the analysis.

Prepare standards by bubbling gas standards into a 500 mL flask filled with deionized (DI) water in a temperature controlled bath at 21 °C. To achieve saturation of the water, use a flow of 12 mL/min for two hours.

Completely fill a 20 mL headspace vial with saturated water and cap immediately without leaving headspace in the vial. This sample is referred to as the bulk sample. Take aliquots were from the saturated bulk sample using a valve syringe. These aliquots are used to build the calibration curve by spiking increasing volumes of the bulk sample into blank DI water samples.

Optimization of saturation times.

5 4 Area

Goal

3 2 1 0 0

1

2

3

4

Hours

Chromatogram of the four gases analyzed (highest calibration point).

Figure 2 1,800

2

Peaks: 1. Methane 2. Ethylene 3. Ethane 4. Propane

Method Setup A method was developed for the Thermo Scientific™ TriPlus™ 300 Headspace Autosampler and the TRACE™ 1310 Gas Chromatograph (Table 1).

5

4 pA

3

Methods The TRACE 1310 GC system is equipped with one Instant Connect Split/Splitless (SSL) injector with a headspace liner (P/N 453A1335) and an Instant Connect Electron Flame Ionization Detector (FID). The column used is a TracePLOT TG-Bond Q 25 m, 0.53 mm (P/N 26004-6120). The gas standards used for calibration may be ordered from Air Liquide. The data are collected and processed using the Thermo Scientific™ Dionex™ Chromeleon™ 7.2 Chromatography Data System (CDS) software.

Results and Discussion The saturation time was optimized by preparing the highest methane calibration standard at 30-minute intervals, while continuously flowing DI water (Figure 1). Other apparatuses and conditions may lead to different results in terms of saturation times. The plot-type column used for this experiment is ideal for effectively retaining and separating compounds such as the gases analysed here. The FID guarantees excellent sensitivity and a wide range of linearity. Chromatograms of the analyzed gas mixture are shown in Figures 2 and 3.

Two calibration curves were built.

The calibration curve for low-level concentrations was suitable for detection of minimum traces of the an-

1 0 –200 0

1

2

3

4

5

6

7

8

8.5

Minutes

In order from left to right, the peaks represent methane, ethylene, ethane, and propane.

Figure 3

Chromatogram of the four gases analyzed (second calibration point, 10 μL added).

10 Peaks:

1. Methane 2. Ethylene 3. Ethane 4. Propane

2 pA

4 3 1

5.5 0

1

2

3 Minutes

4

5

6

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Laboratory Focus June/July 2016

Feature Figure 4

Low-level calibration curves for methane and ethylene.

Figure 6 High-level calibration curve for methane.

alytes in the samples and assessing the sensitivity of the system. Another calibration curve was built for higher concentration intervals, including the saturation level most suitable for screening real samples. Calibration curves were built by adding aliquots of

Table 1

saturated water directly to 10 mL of DI water in 20 mL vials. For low level detection, aliquots of 5, 10, 25, 50, and 100 μL of saturated water were added. For the high level curve, aliquots of 25, 50, and 100 μL and 1, 5, and 10 mL of saturated water were used.

The gas content of each calibration point was calculated from the solubility of the gases at 21 °C, the temperature at which the standards are prepared.

Solubility of the tested gases was: • Methane: 23 mg/L • Ethylene: 149 mg/L • Ethane: 62 mg/L • Propane: 77 mg/L Tables 1 and 2 report the gas concentrations for each calibration point.

The results of the calibration show

Figure 5

Low-level calibration curves for ethane and propane.

Table 3

High-level calibration results for the four gases.

Analyte

r2

Methane

0.998

Ethylene

0.998

Ethane

0.998

Propane

0.995

good performance in terms of linearity with a r2 value above 0.99 for each one of the tested gases (Figures 4–6).

Concentration at the calibration points for the low-level calibration curve.

Level 1 (ppb)

Level 2 (ppb)

Level 3 (ppb)

Level 4 (ppb)

Level 5 (ppb)

Ethylene

12 Level 1 (ppb) 75

23 Level 2 (ppb) 149

57 Level 3 (ppb) 370

115 Level 4 (ppb) 745

230 Level 5 (ppb) 1490

Methane Ethane

12 30

23 62

57 155

115 310

230 620

Ethylene Propane

75 38

149 77

370 192

745 385

1490 770

Ethane

30

62

155

310

620

Propane

38

77

192

385

770

Methane

Table 2

Level 1 Level 2 Level 3 for the Level 4 Level 5 Level 6 Concentration at the calibration points high-level calibration curve. (ppb)

(ppb)

(ppb)

(ppm)

(ppm)

(ppm)

Ethylene

57 Level 1 (ppb) 370

115 Level 2 (ppb) 745

230 Level 3 (ppb) 1490

2.3 Level 4 (ppm) 14.9

11.5 Level 5 (ppm) 74.5

23 Level 6 (ppm) 149

Methane Ethane

57 155

115 310

230 620

2.3 6.2

11.5 31.0

23 62

Ethylene Propane

370 192

745 385

1490 770

14.9 7.7

74.5 38.5

149 77

Ethane

155

310

620

6.2

31.0

62

Propane

192

385

770

7.7

38.5

77

Methane

Conclusion This application is a viable solution for monitoring dissolved gases in water. The analytical procedure and sample preparation are simple and straightforward. Sample conservation and custody are the primary concerns. This method can be used to detect and quantitate both low and high levels of dissolved gases in water samples, making it an excellent option for testing possible impacts of hydraulic fracturing as well as gases in water from surface, non-well stimulated sources.

The 120-position tray of the TriPlus 300 HS autosampler coupled with the exclusive modularity concept of the TRACE 1310 GC offer high throughput capabilities with virtually no downtime for maintenance or switching to other analyses. Altogether, the system is a simple, robust and reliable option for performing this type of analysis in an unattended fashion.

References 1. Sample Preparation and Calculations for Dissolved Gas Analysis in Water Samples Using GC Headspace Equilibration Technique; U.S. Environmental Protection Agency, RSKSOP-175, Revision No. 2, May 2004.

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b y Brig it te Kl ose, Eppendo rf AG

How “Fitting” Pipette Tips Interfere with Assay Results There is more to pipetting than the simple transfer of small and smallest amounts of liquid. The exact and coordinated interaction between pipette and pipette tip is of critical importance. Both components, each on its own as well as together as a system, must meet high quality standards in order to be able to deliver reliable, precise, and reproducible results. With this article, we want to sensitize you to the serious influence pipette tips have on the pipetting result. hen was the last time ISO 8655:2002 recommends the use of pipette and pipette tip by the same manufacturer. epT.I.P.S. are optimally compatible with Eppendorf pipettes.

W

you were upset about non-reproducible results, for example in a real-time PCR? You checked all the variables in your experiment but were still unable to find the cause?

During pipetting (the most frequent step in any experimental process) you always used a pipette with a “fitting” tip – thus a system made up of two components, the precision and accuracy of which you thought you could rely on during the whole workflow. But perhaps you could not?

Purchasing processes with far-reaching consequences

The continuously high quality of the tip is a prerequisite for reproducible results. The image shows an epT.I.P.S.® pipette tip with ergonomically optimized cone geometry, easily recognizable by the droplet-shaped relief elements near the upper rim.

Great attention is paid to the purchase of a new pipette in the laboratory. Its technology must be up-to-date, and handling must be ergonomic.

It should be robust and resistant to chemicals, and of course deliver accurate and precise results. The latter,

however, can only be achieved with the right pipette tip. And this is where contradictions frequently arise: While the pipette is chosen with great care, the selection of the pipette tip is often subject to drastically lower expectations (and budgets), assuming that they are just another consumable.

Especially when purchasing products destined for daily routine, such as pipette tips, many laboratories favour price over quality, unfortunately, without knowledge of the consequences. Many non-reproducible experimental results – and therefore additional costs and expenditure of time – could be avoided if suitable pipette tips were awarded more consideration and selected on the basis of solid expertise.

Study examining pipette tips by 15 manufacturers We have approached this topic by conducting a broadly designed study of pipette tips by 15 global manufacturers (see Application Note at the end of this article). Of course, many more manufacturers exist; however, the tips selected by us are those which in our experience are most frequently found in laboratories, without discriminating between tips by “system providers” of pipettes and pipette tips and “non-system providers”, i.e. manufacturers of plastic labware only.

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Recommendations of ISO 8655:2002 The international standard ISO 8655:2002 recommends the use of pipettes and tips by the same manufacturer because even if the pipette tips by manufacturer A “fit” the pipette by manufacturer B, this does not speak to the precision and accuracy of the pipetting result.

After testing two different volume sizes of pipette tips by all 15 manufacturers on the same pipettes, it became obvious: the pipette tip significantly impacts the result. When using pipette and pipette tip by a system provider, the user can safely assume that errors will not occur, since pipette and tip are calibrated and adjusted by the manufacturer in

form calibration and, if applicable, adjustment of the pipette with the other tips – every time a new lot is used.

accordance with ISO 865 Users can rely on the fact that each lot of the manufactured pipette tips will correspond exactly to this calibration result, i.e. no altered results will be obtained during pipetting. The system manufacturer tightly coordinates his production of pipettes and tips with respect to production tolerances and thus ensures that the system operates within specifications at all times.

This is not the case for non-system manufacturers; they do not have system specifications. If a pipette by one manufacturer is used with a pipette tip by a different manufacturer, neither of them will take responsibility for the precision and accuracy of the system.

In this case the user is forced to per-

Tip design also plays a role During our studies we found out that the tip design, too, has a large impact on the pipetting result. Those who pipet a lot are often familiar with pipette tips by different manufacturers and know that many different shapes of pipette tips exist.

Some are longer and slimmer than others. There is the “rocket shape”, as well as the regular conical pipette tip.

Such different shapes influence the pipetting process. The longer the tip, with otherwise identical inner dimensions, the higher the air-cushion inside the tip. Since pipettes are always

feature adjusted to a defined air-cushion inside the pipette, a change in air-cushion, especially for larger nominal volumes, will impact accuracy.

Often the user is not aware of the consequences the final result of their work will suffer as a result of a careless choice of pipette tips. This may be one reason why scientific publications rarely, if ever, list the complete name, manufacturer, and lot number of the tips used. Errors caused by the pipetting system cannot be traced, and reproduction of the data by other scientific groups is no longer possible.

Don’t let it get this far and read more on this topic in the Application Note below.

b y M u riel Art, Vincent Duf ey, Ioa n Gligor , Ep p end or f Ap p l ic atio n Techno lo gies S.A ., Namur , Belgium Ulrike G a st, Ronja Kuba s ch, Eppendo rf Ag, Hamburg, Germany

Application Note: The Tip of the Iceberg: How Pipette Tips Influence Results.

Part 1: Tip Fit Is Not All Users Should Look for Abstract The fact that a tip fits onto a pipette cone does not say anything about the performance of the pipetting system comprising the components “Pipette and Tip”. We performed a study including standard tips from 15 different manufacturers in order to investigate the tip-related influence on the pipetting result. The study results showed a dramatic influence of the tip on the pipetting accuracy.

Figure 1

The standard ISO 8655:2002 [1] recommends using pipette and tips from the same supplier. Our study results emphasize the validity of this recommendation and the need of calibration/pipette adjustment if other tips than recommended by pipette suppliers are to be used.

Introduction Within the scientific community, a rising number of published experi-

ments cannot be reproduced by other groups. In general, plastics are not taken seriously leading to problems with analysis results caused by e.g. leachables or incorrect pipetting volumes. This may lead to results not being reproducible if performed by other groups using other consumables.

Some problems with pipette tips are obvious, e.g. the need to push tips with force onto the pipette cone in order to achieve efficient tip fit.

Other problems often remain unrecognized like decreased pipette accuracy when using other tips than recommended by the pipette supplier.

The ISO 8655-2:2002 standard 1 defines pipette and tip as a system, which requires extra calibration for the use of other manufacturers’ tips.

But why does this standard put so much focus on a product that is to be discarded after usage?

This article answers this question.

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June/July 2016 Laboratory Focus www.laboratoryfocus.ca

Feature

Figure 2

It shows the influence of tips on the pipetting result explaining the main tip-related impact factors.

Material and methods General material Eppendorf Xplorer® plus 50 –1,000 μL and 0.5 –10 μL were used. Racked 10 μL and 1,000 μL standard tips of Eppendorf and 14 other manufacturers have been tested. Exceptions: Manufacturer H did not offer racked 10 μL standard tips, manufacturers K and N offered only 1,250 μL tips for 1,000 μL pipettes.

Calibration by gravimetric method The performance of the system “Pipette and Tip” was determined by calibration according to [1]. Environmental conditions were set according to requirements [1].

Calibration was performed using analytical balance Model XP26PC (Mettler- Toledo®) at 100 per cent nominal volume and 10 per cent nominal volume. Two series of 10 pipettings were performed. Systematic error and random error were determined for each series of 10 measurements and compared to specifications [1] and [2].

For further information please refer to [3].

Results and discussion While being perfectly within error limits with Eppendorf tips, we found the system “Pipette and Tip” to be out of specifications when using other manufacturers’ tips.

As shown in Fig. 1 and 2, the allowed systematic error was exceeded with four manufacturers’ tips at a volume of 1,000 μL and with 5 manufacturers’ tips at 1 μL test volume. These tips with 1,000 μL test volume exceeded not only the manufacturer specifications but also the wider limits for systematic error as stated by the ISO 8655:2002 standard [1]. The random error was noticeably increased but stayed within allowed tolerances.

When comparing the calibration results with the outcome of dimensional measurements, it becomes clear that with 1,000 μL the biggest impact factor is the air-cushion size: Those tips that produced error limits beyond the pipette specifications were longer although they had a similar inner diameter [3].

Larger tips offer a larger air-cushion. Since air-cushion pipettes are adjusted to a certain air-cushion size, an increase in air-cushion volume has a negative impact on the pipetting result, especially with nominal volume.

The calibration results have been found to be independent of the pipette manufacturer and were reconfirmed by calibrating with another manufacturer’s pipette (data not shown).

In contrast to 1,000 μL tips, the impact of “air-cushion size” does not influence the 10 μL tips to an important degree. With this tip model, other influencing factors such as geometry and quality of tip orifice are more important. These influencing factors are discussed in more detail in [3] Part

2 of this application note which will appear in Eppendorf’s publication BioNews.

The results are conclusive that precaution is needed if tips with a design differing from the tip recommended by the pipette supplier are to be used. Elongated tips are an example. Within the operating manual, Eppendorf advises the user to adjust the pipette if such tips are to be utilized. In the case of manual pipettes, this can be done by user adjustment.

More conveniently, with electronic Xplorer pipettes, this is performed by just choosing the tip from the options menu. Most non-system tip providers inform about tip fit on different pipettes. However, the fact that a tip physically fits onto a pipette cone does not say anything about the system’s performance. If other manufacturers’ tips are to be used, the only user’s chance to learn about inaccurate pipetting results is by calibration.

This is supported by statements from the standard [1] which generally recommends using pipette and tip of one manufacturer. In case this is impossible, the user is required to calibrate first with the tips recommended by the pipette manufacturer (“conformity testing” to ensure the system is working correctly), and secondly to calibrate with other manufacturers’ tips [1].

Conclusion That a tip physically fits onto a pipette cone does not mean that the

system performs within the manufacturer’s specification. We have shown for 1,000 μL and 10 μL tips that the system’s performance can be heavily influenced by the tip.

Manufacturer-wise pipettes are supplied adjusted to a certain aircushion size. The tip design, however, directly influences the air-cushion size. Especially with bigger volumes such as 1,000 μL, this happens to a degree affecting the system’s accuracy. With small volumes, other impacting factors come into play which will be discussed in more detail in Part 2 of this application note which will be available shortly.

References: 1. DIN EN ISO 8655:2002, parts 1, 2, 6. Piston-operated volumetric apparatus. Beuth-Verlag, Berlin, Germany.

2. Eppendorf Xplorer® plus operating manual. www.eppendorf. com/manuals 3. Application Note No. 354: The tip of the iceberg: How pipette tips influence results. www.eppendorf.com/application

To see this story online visit http://www. laboratoryfocus.ca/ how-fitting-pipette-tips-interfere-with-assay-results/

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Laboratory Focus June/July 2016

VISCOMETER RheoSense Inc. introduces the VROC® initium, an automatic viscometer for viscosity characterization. Equipped with automatic sample loading and sample cleaning, VROC® initium measures absolute viscosity as a function of shear rate across a wide temperature range.

This allows unique viscosity fingerprinting of the samples. Samples in 96 well plates as well as 40 vial racks can be tested automatically with the intuitive software. It can measure viscosity for sample volumes as little as 10 microlitres, making it suitable for early stage development of protein therapeutics . Automatic intrinsic viscosity measurements provide size of molecules in various formulations and in various conditions which help shorten development time. With the extreme repeatability and small footprint, the VROC® initium is the workhorse for both research and routine viscosity measurements.

Web: www.rheosense.com

CAMERAS ZEISS has introduced two new digital microscope cameras, the ZEISS Axiocam 702 mono and ZEISS Axiocam 512 color, to its current portfolio.

With ZEISS Axiocam 702 mono, ZEISS for the first time introduces a microscope camera with a scientific CMOS sensor. Users benefit from low read noise, excellent low light sensitivity and high speed for live cell imaging and acquisition of fast processes. ZEISS Axiocam 702 mono features a 1/1.2″ (13.3 mm diagonal) sensor with a resolution of 2.3 megapixels, making it an attractive entry into the world of scientific CMOS imaging. ZEISS Axiocam 512 color allows acquisition of large sample areas in one high resolution, true color image. The 12 megapixel CCD sensor with a size of 1″ (16 mm diagonal) delivers an excellent live image and high acquisition speed. ZEISS Axiocam 512 color in combination with low magnification and zoom objectives offers optimal resolution for imaging of large specimens without stitching. Both new camera models feature high-speed USB 3.0 connections and active thermoelectric cooling.

Web: www.zeiss.com

DNA EXTRACTION KIT The NextPrep-Mag™ Urine cfDNA Isolation Kit from Bioo Scientific is an automation-friendly kit designed to extract cell free DNA from urine, using a magnetic bead format. The rapid procedure can be completed in only 30 minutes and does not require a vacuum manifold. Using the kit, sufficient cfDNA is recovered from urine sample volume ranging from less than 1 mL to more than 20 mL, to allow robust construction of libraries for targeted or whole genome sequencing. The cfDNA isolated using this kit can be used for applications including studies of circulating fetal DNA and cancer diagnostic-related liquid biopsy studies.

Web: www.biooscientific.com

New Products PIPETTES Unlike traditional pipettes which utilize a single rotating plunger to set volumes, the EVOLVE manual pipette from INTEGRA features three adjustable dials for setting each individual volume digit.

Simply depress and twist the plunger to unlock the volume dials. Once unlocked, freely adjust the three dials to rapidly set the desired volume. This new approach allows users to set volumes more than ten times faster. The pipettes come in single, eight and twelve channel formats, covering a volume range of 0.2 to 5,000 µl. They are optimized for use with INTEGRA’s wide range of GripTips pipette tips. The GripTips simply snap into place with minimal tip loading effort, providing a secure connection. Additionally, the new INTEGRA linear stand can accommodate up to thirteen EVOLVE pipettes. It can also be converted into a charging stand to charge any INTEGRA electronic pipettes.

Web: www.integra-biosciences.com

IMAGING SYSTEM Molecular Devices LLC announces the launch of its ImageXpress® Micro 4 HighContent Imaging System.

Additionally, its latest version of MetaXpress® HighContent Image Acquisition and Analysis Software is now capable of 3D image analysis.

For scientists conducting basic research or high-throughput screening, the ImageXpress System is an integrated and scalable toolset for optimal 3D image acquisition and analysis. With the new ImageXpress system and MetaXpress 3D software toolkit users can directly access a smooth and integrated workflow, eliminating the need for third-party tools and time-consuming bottlenecks for acquisition and analysis of complex 3D datasets. Its modular configuration allows researchers to customize the system to fit individual needs, now including the ability to add confocal capability with the AgileOptix™ spinning disc confocal module without disruption to existing workflows.

Web: http://go.moleculardevices.com/ixm-4

SAMPLE PREPARATION Seward Ltd. has introduced the all new Seward Stomacher® 400 EVO laboratory blender, together with Stomacher® 400 ECO sample preparation bags.

The Stomacher 400 EVO has been engineered and designed for easy use and silent operation. Laboratories can also save on consumables by using the blender with the new Stomacher ECO bags. These use significantly less plastic, saving waste and making them both economical and eco-friendly. Its unique, one-touch bag loading system means operators just need to place the bag in the loading slot, and by pushing the start button, the bag is sealed and blending begins. Together with the ‘no-fuss’ programmable screen display, operators can process samples quickly and efficiently. The Seward Stomacher 400 EVO’s rubber base, combined with a host of design and engineering developments, also helps to eliminate bench movements. A removable drip tray means that any spillages during the day can be quickly cleaned, and for thorough cleaning, the paddle chamber can be easily opened and the paddles removed, making the complete chamber accessible.

Web: www.seward.co.uk.

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New Products HOT PLATES Torrey Pines Scientific has released its new EchoTherm™ Model HS65 programmable digital stirring hot plate for use in chemical, pharmaceutical, environmental, biochemical and other laboratories. With the EchoTherm™ Model HS65, programming is done through the front panel membrane switch and full-functioned custom liquid crystal display. It comes with five-stirring positions allowing for reproducible, accurate, hands-off sample preparation. The unit can store 10 programs in memory of as many as 10 steps each where each step is a temperature, temperature ramp rate (if wanted), stirring speed and time. Each program can be made to repeat itself automatically from 1 to 98 times or infinitely. All programs are stored electronically. Each stirring position can be set to the same speed or to individual speeds.

Non-programmed operation can be done as well. Heater plates are 12″ (30.5 cm) x 12″ (30.5 cm) white, solid ceramic glass for excellent chemical resistance and quick heating. The plate surface can be heated from ambient to 400°C. Plate surface temperature or solution temperature is controlled directly to 1°C of the target. Accuracy is 1% of the reading using platinum RTD circuitry. The units are supplied with immersion probe and temperature calibration certificate traceable to NIST. Stirring speed is 100 to 1500 rpm and controlled by optical coupler to 10 rpm.

Count down timer with alarm, user settable Auto-Off, and RS232 I/O port are standard with the unit. The Model HS65 is available in 100, 115, and 230 VAC, 50/60 Hz models. All units are UL, CSA, and CE certified.

Web: www.torreypinesscientific.com

BIOCHEMISTRY ANALYSIS Beckman Coulter Life Sciences has unveiled its new low sample volume bioanalyte analyzer, the ViCELL MetaFLEX*. Built for micro to large-scale cell culture applications, the biochemistry analyzer delivers fast, accurate bioanalytical analysis (measuring pH, pO2, pCO2, glucose, lactate, electrolytes and more parameters). The compact Vi-CELL MetaFLEX is designed to be operational more than 22 hours a day, with a turnaround time of less than 60 seconds. The magic is in the sensor cassette. The sensor is based on thick-film technology – several layers of ceramic slabs, microelectrodes and membranes; miniaturization of conventional electrodes. The result is a shorter measuring time, and smaller sample size. The instrument is able to store up to 2,000 results on-board, with an automated sampling arm so that samples can be introduced from a syringe, tube, microfuge or Vi-CELL sample cup. Testing is quick and intuitive. The only consumables required are the sensor cassette and solution pack. The solution pack includes reagents, wash solutions, QC controls and waste collection. Inserting a new cassette and solution pack is the only maintenance needed. The Vi-CELL MetaFLEX’s quality management system automatically measures on three dedicated QC solutions, detecting and correcting any failures, and locking out any parameter that fails QC.

Web: beckman.com

June/July 2016 Laboratory Focus www.laboratoryfocus.ca

CELL IMAGING BioTek Instruments announces the launch of its Lionheart™ FX Automated Live Cell Imager with Augmented Microscopy™. Lionheart FX is optimized for kinetic live cell imaging, with up to 100x air and oil immersion magnification in a variety of slides, dishes, microplates and flasks. Imaging channels include brightfield, colour brightfield, phase contrast and fluorescence, with both image-based and laser autofocus. A unique environmental control cover provides convenience beyond that of typical digital microscopes, allowing for incubation up to 40º C and gas control, while the optional humidity chamber and dual reagent injectors enhance environmental support for live cell imaging workflows. Gen5™ 3.0 Software provides automated image capture and analysis, plus annotation and movie maker functions, offering ease and simplicity across a broad range of live and fixed cell applications. Augmented Microscopy is the combination of all of these features in one compact system. Lionheart FX is ideal for live and fixed cell applications including kinetic live cell assays, 3D spheroid imaging, translocation and colocalization studies, cell migration and invasion, proliferation, viability, toxicity and many more.

Web: www.biotek.com.

ASSAYS Aushon BioSystems has released its new Ciraplex™ ULTRA Ultrasensitive Assays with multiplexing capabilities.

The assays with femtogram/ml (fg/ml) levels of detection address multiple biomarkers in a wide range of therapeutic areas. These kits offer enhanced sensitivity and an extended dynamic range providing quantification below standard detection limits. As well, intuitive Cira™ software streamlines the data management process providing exceptional data control.

Web: www.aushon.com

CELL CULTURE Nova Biomedical has added the BioProfile FLEX 2 to its BioProfile line of cell culture chemistry analyzers. BioProfile FLEX 2 incorporates Nova’s maintenance-free MicroSensor™ Card technology that eliminates sensor maintenance and maximizes workflow efficiency. BioProfile FLEX 2’s very small sample volume allows for automated cell culture analysis for small volume culture systems, such as the Sartorius Ambr® microbioreactor systems.

BioProfile FLEX 2’s innovative design provides comprehensive analysis of 16 key cell culture chemistries including Gluc, Lac, Gln, Glu, NH4+, Na+, K+, Ca++, pH, pCO2, pO2, Osmolality by freezing point depression, and Total and Viable Cell Density and Viability by trypan blue dye exclusion method. The full panel of tests can be run in under four minutes with just 265 microliters of sample, and users can select from manual sampling or two modes for batch sampling: 96-well plates or the builtin 24-position tray. Designed for small volume, early stage cell culture development through GMP manufacturing, BioProfile FLEX 2 provides 21CFR Part 11 compliance, OPC connectivity, full networking functionality, and support with all validation needs.

Web: www.novabiomedical.com

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Laboratory Focus June/July 2016 www.laboratoryfocus.ca

Engaging Canada’s young scientists On May 6, 2016, as part of a special kick off to ScienceOdyssey, a 10-day national celebration of science-based activities and events across Canada, Canada’s federal government announced it was committing $4.8 million in funding towards science education initiatives at 43 different organizations across the country.

The announcement was made at the Canadian Association of Science Centre’s annual conference in Vancouver, BC, where Canada’s Minister of Science Kirsty Duncan was on hand to unveil the new PromoScience Program at the Natural Science and Engineering Research Council of Canada (NSERC). Minister Duncan discussed the merits of the program, and its mandate to enable science outreach across Canada. The program hopes to fulfill its mandate through the support of science centres, day camps, after-school programs, science outreach organizations, networks and more. Essentially, it hopes to encourage the engagement of youth in the science and engineering fields.

The Minister also used the PromoScience Program launch as an opportunity to reemphasize her government’s commitment to promoting more openness and transparency when it comes to the communication of science and evidence-based policymaking. There was also some further discussion around the previously noted establishment of a Chief Science Officer for Canada.

“I have examined how similar positions, often called a chief science adviser, work in other countries such as the United Kingdom, New Zealand, the United States and Israel. My survey of international models will help create a position that is modern and yet tailor-made to suit Canada,” she said.

As for how the 10 day science celebration played out, activities included science in the streets, visits to labs, science fairs, talks, conferences, school field trips, encounters with researchers and scientists, special exhibits at museums and science centres, and much more.

The celebration of youth science continued later in May with the CanadaWide Science Festival. In all, nearly $1 million in cash awards and scholarships were awarded to some of Canada’s most promising young scientists from Grades 7 to 12. Among the winners were Kayley Ting from Richmond Hill, ON who won the Best in Fair and Intermediate Platinum award for her project: Analysis of Electrodermal Activity to Quantify Stress Levels in Autism. The Senior Platinum award went to Katherine Teeter from Markdale, ON, for her project: Synthetic Limpet Teeth for Improved Joint Performance, and finally the Junior Platinum award went to Sophie Hoye Pacholek from Calgary, AB, for her project: The Genius Genus: Aspen Adaptation.

Likewise, as per our cover story, Iveta Demirova from New Westminster Secondary School in New Westminster, BC took top honours at the national final of the prestigious Sanofi Biogenius Canada (SBC) competition in Ottawa.

The 16-year-old, grade 11 student, was chosen by the judges for her research project exploring the development of a novel HIV-1 therapy. The results of Iveta’s research project, completed with the support of mentor Dr. Ralph Pantophlet of Simon Fraser University, could offer numerous advantages to those living with HIV, which remains one of the world’s leading infectious diseases.

She will now progress to the 2016 International BioGENEius Challenge in San Francisco in June, where she will submit her work to a panel of pre-eminent international scientists. We wish her, and her fellow Canadian competitors luck, and we’re excited to see them shine on the world stage!

app review The Elements A Visual Exploration From Touch Press for iOS/iPad and iPhone http://apps.theodoregray.com/the-elements/

Designed for chemistry students, and pretty much anyone interested in science, this app based on Theodore Gray’s The Elements brings users the periodic table like they’ve never seen it before. Each element has two pages devoted to it, including a 3D virtual reality (VR) depiction of the element, either in its natural form or some product made from it, along with its chemical symbol, atomic number. Users can even view individual elements via an animation set to a theme song. For recently discovered elements, an image of the person it’s named for is shown instead, but the site does include a breakdown for these elements as well. In general, users can rotate and manipulate any of the 500+ objects with a simple sideways swipe or by touching the illustration. Along with extensive background detail for each element, the app also embeds real-time computational data from Wolfram Alpha. Our verdict, this app is a both entertaining and educational, and worth the asking price.

PubMedOn Tap

By ReferencesOnTap for iOS-based mobile devices https://itunes.apple.com/us/app/pubmed-on-tap/ id301316540?mt=8 With PubMed On Tap users can search PubMed and retrieve articles directly from their phones or tablets. According to the tool’s iTunes page, users can also “search personal reference libraries, organize references in groups (static and smart), and email references from their device to themselves or others.” The Advanced Search screen lets you pick your fields without having to know field code abbreviations, which is probably more helpful for most users. Additionally, two concepts can be searched at a time. Numerous search options from PubMed are available, including publication type, date, subset (which includes systematic review and core clinical journal filters), and gender. Once you view a PDF, it downloads to the app and becomes accessible through the Authors and Attachments Smart Groups. The makers of this app offer two versions: PubMed on Tap and the free PubMed on Tap Lite. The two are essentially the same, except that the Lite version includes ads, displays only 20 references per online search, and allows you to save only 50 references. We recommend you try it first to see if this app meets your needs. Unfortunately the app can’t be used by Android users, however, they have PubMed Mobile as their option.

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Laboratory Focus June/July 2016

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48th Annual Canadian Mineral Analysts (CMA) Conference and Exhibition Venue: Sudbury, ON Tel. 705-670-5643 Fax. 705-670-3047 June 12-15 E-mail: ed.debicki@ontario.ca Web : 10:01 www.2016cma.com Canadian Society for Microbiologists Annual RC_lab_new:Layout 1 12/19/2012 AM Page 1 Conference September 13-15 Venue: Toronto, ON Biopharm America Tel: 613-421-7229 Venue : Boston, MA Email: info@csm-scm.org Web : http://www.ebdgroup.com/bpa/index.php Web: www.csm-scm.org/english/conf_upcoming.asp CALAS Annual Symposium Venue: Toronto, ON Tel: 416-593-0268 Web: www.calas-acsal.org/symposium

SOCRA Conference Venue: Montreal, QC Email: registration@socra.org Web: www.socra.org

OCTOBER October 16-19 Canadian Chemical Engineering Conference Venue : Quebec City, QC Tel : 613-232-6252 Web : www.csche2016.ca

June 14 MedEdge Summit Venue: Richmond Hill, ON Livestream: https://livestream.com/ accounts/1705310/events/4100246 Web: www.businessrichmondhill.ca/MedEdge/

SRC103

June 14-16 Canadian Biosafety Symposium Venue: Montreal, QC Tel: 306-966-8496 Web: http://biosafety.icid.com

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