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February/March 2016 Volume 20, Number 1

Improve Nucleic Acid Extraction

Protein Quality Control in SPR and BLI

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

McGill astrophysicist wins Herzberg Gold Medal galaxy. She is also the first woman to win the award. In addition to the Herzberg Canada Gold Medal, NSERC also awarded prizes to other researchers for outstanding university-industry partnerships, ground-breaking discoveries, excellence in multidisciplinary research, and fellowships to enhance the career development of outstanding and highly promising scientists and engineers. These award recipients include: NSERC John C. Polanyi Award: Barbara Sherwood Lollar, University of Toronto The Brockhouse Canada Prize for Interdisciplinary Research in Science and Engineering: Shana Kelley, University of Toronto, and Edward H. Sargent, University of Toronto

His Excellency presents the 2016 Gerhard Herzberg Canada Gold Medal for Science and Engineering to Victoria Kaspi. Credit: MCpl Vincent Carbonneau, Rideau Hall, OSGG.

Ottawa, ON – McGill University astrophysicist Dr. Victoria M. Kaspi has won the Gerhard Herzberg Canada Gold Medal for Science and Engineering, awarded by the Natural Sciences and Engineering Research Council of Canada (NSERC).

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The prize, which comes with a $1 million research grant, has been awarded annually since 1991 to recognize “sustained excellence and overall influence” of research conducted in Canada. Dr. Kaspi is one of the world’s

leading experts on neutron stars, the ancient remnants of the most massive stars in the Milky Way. Using the largest and most powerful radio and X-ray telescopes in the world, Dr. Kaspi studies the physical behaviour of these types of stars, pulsars and magnetars. Her research group has had major impacts in the field of astrophysics, including unique tests confirming Einstein’s long-held theory of general relativity and discovering the fastest rotating star. Her team’s 2002 landmark discovery of powerful X-ray bursts from an enigmatic class of star essentially doubled the number of known magnetars in our

NSERC Gilles Brassard Doctoral Prize for Interdisciplinary Research: Yasser Gidi, McGill University E.W.R. Steacie Memorial Fellowships: Elena M. Bennett, McGill University, Curtis P. Berlinguette, The University of British Columbia, Zhongwei Chen, University of Waterloo, David Sinton, University of Toronto, Mark Vellend, Université de Sherbrooke, and Stephen I. Wright, University of Toronto Synergy Awards for Innovation: Category 1: Small- and Mediumsized Companies: Emil M. Petriu, University of Ottawa, Industrial partner: Rami Abielmona, Larus Technologies Corp. Category 2: Large Companies: J. David Miller, Carleton University, Industrial partner: Greg Adams, J.D. Irving, Limited Continued on page 3

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GOVERNMENT OF CANADA INVESTS IN RESEARCH COMMERCIALIZATION Charlottetown, PEI – The federal government says it is investing $29 million towards the establishment of two new commercialization centres.

Minister of Agriculture and Agri-food Lawrence MacAulay and Prince Edward Island Premier Wade MacLauchlan. The first commercialization centre, Natural Products Canada (NPC), will receive $14 million over five years, in sup-

The announcement was made by Canada’s Minister of Innovation, Science and Economic Development Navdeep Bains, alongside the Canadian

port of its work to establish Canada as a global leader in the development and marketing of natural products. The second, the Centre for Commercialization of Cancer

Continued on page 3

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Continued from page 2 Immunotherapy (C3i) will receive funding of $15 million over five years to develop, translate and commercialize cancer immunotherapies. The two recipients were chosen from the most recent competition in the Centres of Excellence for Commercialization and Research (CECR) program. The CECRs match clusters of research expertise with the business community, facilitating the development of products and technologies

at a stage in the commercialization process where it is otherwise difficult to attract private-sector investment. The NPC, headquartered in Charlottetown, includes key partners in PEI (PEI BioAlliance as NPC Atlantic), Saskatchewan (AgWest Bio as NPC West), Ontario (Ontario Bioscience Innovation Organization as NPC Ontario), and Québec (Institute for Nutrition and Functional Foods as NPC Québec). The federal contribution will be matched

by over $10 million from industry and other sources, for total funding of over $24 million over the next five years. C3i, which is based in Montréal, will accelerate access to innovative cancer immunotherapies for patients. Operating out of the Hôpital Maisonneuve-Rosemont’s Research Centre, C3i will be a one-stop shop for the development, translation and commercialization of ground-breaking cancer treatments.

PROOF Centre collaboration focuses on prognostic biomarkers for chronic kidney disease

Vancouver, BC – The Centre of Excellence for the Prevention of Organ Failure (PROOF Centre) has launched a new partnership with AstraZeneca and the Canadian Study of Prediction of Death, Dialysis and Interim Cardiovascular Events (CanPREDDICT) to develop a blood test to predict the rate of disease progression in patients with

Continued from page 1 Category 3: Two or More Companies: Jean Caron, Université Laval, Industrial partners: Jean-Paul Guérin, Maraîchers J.P.L. Guérin et fils inc., Daniel Malenfant, Vert Nature inc., Denys Van Winden, Production horticole Van Winden inc., Jean-Bernard Van Winden, Les fermes Hotte et Van Winden inc., and Stéphane Van Winden, Delfland inc. Category 4: Colleges: Neil Cooke, Red River College; Ray Hoemsen, Red River College; Jose Rizalino M. Delos Reyes, Red River College; Shokry Rashwan, Red River College; Rob Spewak, Red River College; Industrial partner, Lloyd Kuczek, Manitoba Hydro To see this story online visit http://www.laboratoryfocus.ca/ mcgill-astrophysicist-wins-herzberggold-medal/

chronic kidney disease (CKD). For the discovery phase of the project, PROOF Centre and AstraZeneca researchers will leverage the large cohort and rich clinical data available from the CanPREDDICT study. The collective team of clinical, computational, technical and biological experts will use the

PROOF Centre’s computationallydriven omics biomarker development pipeline to identify genomic and proteomic biomarkers that can discriminate patients who have rapid versus slow CKD progression. The PROOF Centre will support the analysis process from front-end experimental design to downstream statistical and biological analysis. “Biomarkers to better predict disease progression will be a valuable tool to provide more personalized care for patients suffering from CKD,” says Dr. Adeera Levin, CanPREDDICT lead investigator and head of UBC Division of Nephrology. “If developed as a blood test that corresponds to different patient trajectories, it will offer clinicians a better way of knowing which patients need more intensive medical attention and management.” Dr. Levin adds that a prognostic test will allow clinicians to more aggressively manage CKD in patients expected to rapidly decline, potentially delaying end-stage renal disease. Patients whose kidney function is pre-

The CECR program currently funds 23 centres, working in areas that include information and communications technologies, health, natural resources and energy. Created in 2007, the program invests $30 million per year in Canadian innovation. To see this story online visit http://www.laboratoryfocus.ca/ government-of-canada-invests-inresearch-commercialization/ dicted to stay stable or improve can be monitored less frequently. Such a test will also be extremely valuable for the research and drug development community. “Chronic kidney disease is a strategically important research area for AstraZeneca, knowing which patients with this disease are likely to face rapid decline and which ones are not, is a major hurdle in clinical trials for new therapeutics,” Dr. Peter Greasley, director clinical research at AstraZeneca explains. “More than 80 per cent of AstraZeneca’s portfolio across small and large molecules has a personalized healthcare approach and biomarkers identified in this collaboration will enable patient stratification that in turn may reduce the time, number of patients, and costs needed for such trials.” CKD is a major global public health issue. One in 10 adults have some degree of kidney function decline, and CKD is estimated to affect three million Canadians. To see this story online visit http://www.laboratoryfocus.ca/ proof-centre-collaboration-focuseson-prognostic-biomarkers-for-chronic-kidney-disease/

Comet Biorefining to build first commercial biomass sugar plant in Sarnia, Ontario London, ON – Comet Biorefining, Inc. has chosen the TransAlta Energy Park in Sarnia, ON as the location of its commercial-scale biomassderived sugar facility. The 60 million pounds per year plant will come online in 2018 producing dextrose sugar from locally-sourced corn stover and wheat straw. Corn stover consists of residues left in the field after harvest including stalks, leaves, husks and cobs. “Construction of this first-of-a-kind plant represents a key step towards the large-scale commercialization of our cellulosic sugar business,” said Andrew Richard, CEO of Comet. “It highlights the important role our technology plays in the value chain, helping to drive the bioeconomy and

reduce greenhouse gas emissions.” Using its proprietary patented process, Comet converts non-food agricultural and forest residues into high-purity dextrose sugars that will be transformed into bio-based products including organic acids, amino acids and bioplastics. These low-carbon bio-based products replace traditional petroleum-based materials, reduce greenhouse gas emissions and help contribute to Canada’s efforts on climate change. Comet dextrose is costand performance-competitive with commercial dextrose sugars, the benchmark raw material for today’s biochemical production. Comet chose to locate in Sarnia by working together with Bioindustrial Innovation Canada (BIC), the

Ontario Federation of Agriculture (OFA) and an Ontario farmers’ cooperative on a project to attract sustainable technology providers to the region and to meet increasing demand from chemical suppliers and consumers for low-carbon products. “Comet’s cellulosic sugar technology was one of the clean sustainable technologies recommended, with the best fit for the region and an excellent opportunity to accelerate the growth of the bioeconomy in rural Ontario”, noted Dr. Murray McLaughlin, executive director, BIC. To see this story online visit http://www.laboratoryfocus.ca/ comet-biorefining-to-build-firstcommercial-biomass-sugar-plantin-sarnia-ontario/

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British Columbia backs Genome BC with big funding round

Vancouver, BC – Cutting-edge health research projects at Genome BC are getting a big boost with the B.C. government kicking in $34 million to support genomics research in the province.

The funding will support both new and ongoing projects at Genome BC, including cancer projects in hereditary and childhood cancer; pharmacogenomics research into adverse drug interactions; infectious disease projects

in areas such HIV, tuberculosis, Hepatitis C, and Avian flu; and rare diseases. “The research undertaken by Genome BC has a real impact for British Columbians,” said B.C. Premier Christy Clark. “From genome sequencing to quickly diagnose newborns with genetic disorders, to implementing genome testing to save people with lung cancer, HIV and other serious illnesses, our partnership with Genome BC has increased testing efficiency and helped save lives.” One of the projects to be supported is RAPIDOMICS, which uses genome sequencing to identify genetic disorders in newborns at BC Women’s Hospital. Currently, genetic disorders like neurological problems, acute metabolic collapse, and malformations are a leading cause of infant mortality and make up a large percentage of the patients in B.C. neonatal intensive care units (NICU). The genome sequencing under RAPIDOMICS could significantly speed up the time to get an accurate and rapid diagnosis for patients with these types of disorders.

“Rapid exome sequencing in the NICU could replace hundreds of different tests that are currently in use, and greatly enhance support for families at this very difficult time,” said Dr. Alan Winter, president & CEO of Genome BC. “Genome BC is in the business of solving problems impacting British Columbians. This project is an example of genomics enabling the precision and timeliness of patient diagnosis and management as well as reducing costs for the health care system.” The $400,000 RAPIDOMICS project aims to introduce the use of rapid exome sequencing in the NICU, a technique for sequencing the proteincoding genes in a genome (known as the exome). The pilot will include exome sequencing of 25 trios (baby plus both parents). Currently, rapid exome sequencing is not routinely available in B.C. The pilot project will help evaluate whether it should be expanded to serve extremely sick babies anywhere in British Columbia. To see this story online visit http://www.laboratoryfocus.ca/ british-columbia-backs-genome-bcwith-big-funding-round/

Therapeutic approach offers new hope for treatment of multiple myeloma Montréal, QC – A new therapeutic approach tested by a team from Maisonneuve-Rosemont Hospital (CIUSSS-EST, Montréal) and the University of Montréal has shown promising results for the treatment of multiple myeloma. Multiple myeloma is a cancer of the bone marrow currently considered incurable with conventional chemotherapy and for which the average life expectancy is about six or seven years. The study was led by Dr. Jean Roy, a haematologist at the Maisonneuve-Rosemont Hospital and professor at the University of Montréal, between 2001 and 2010 with 92 newly diagnosed patients. Using an approach developed at the hospital, consisting of an autograft to reduce tumour mass followed by a family allograft three to four months later to clean the bone marrow of myeloma cells with immune cells from a family donor (immunotherapy), the study resulted in a total cure rate of 41 per cent, a record level using this strategy. Moreover, patients in complete remission six months

after the allograft had a relapse-free survival rate of 60 per cent. Overall, the autograft strategy followed by allograft resulted in relapse-free survival rates of 20 to 25 per cent in the long term. “In many hospitals, doctors have abandoned the use of allografts for multiple myeloma due to the risk of toxicity and relapse,” said Dr. Roy. “Our results, on the other hand, have led us to offer the treatment to more patients, especially younger patients and those with poorer prognoses.” Moreover, in the same study, the mortality rate associated with this treatment over ten years was 10 per cent, an extremely low rate following a family donor allograft. And while 50 per cent of patients experienced a relapse of their myeloma, the subsequent treatment showed marked efficacy: 50 per cent of these patients were alive five years after the relapse. Based on these results, a new study aiming to reduce rates of relapse and complications is currently underway at Maisonneuve-Rosemont Hospital, with still preliminary but very encouraging results.

The results of this research were published in the specialized journal Bone Marrow Transplantation. The study was supported by the University of Montréal’s William Brock Fund.

To see this story online visit http://www.laboratoryfocus.ca/ therapeutic-approach-offers-newhope-for-treatment-of-multiplemyeloma/

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Dr. Brian H. Rowe, a University of Alberta clinician-scientist, is the new scientific director of CIHR’s Institute of Circulatory and Respiratory Health. Dr. Rowe is a professor of Emergency Medicine and an adjunct member of the School of Public Health at the University of Alberta in Edmonton, a practicing emergency physician and a Tier I Canada Research Chair in Evidence-based Emergency Medicine. He is also the scientific director of

Dr. Brian H. Rowe

the Alberta Health Services’ Emergency Strategic Clinical Network, as well as the former Associate Dean (Clinical Research) and co-director of the Evidence-based Practice Centre in the Faculty of Medicine & Dentistry at the University of Alberta. He obtained his medical degree and received training in emergency medicine at the University of Ottawa; he completed his graduate research training at McMaster University. He is the author of over 440 peer-reviewed publications and 30 book chapters. Xenon Pharmaceuticals Inc. has hired James R. Empfield as its new senior vice president, Drug Discovery. Reporting to Dr. Simon Pimstone, president and CEO, Dr. Empfield will be responsible for leading drug discovery chemistry, manufacturing, and other preclinical activities that support the growth of Xenon’s discovery and development pipeline. Prior to joining Xenon, Dr. Empfield served as vice president, Drug Discovery and Chemistry; co-head of Research, Boston at Vertex Pharmaceuticals Inc. from 2011 until August 2015.

Appointments

From 2006 to 2011, he was director, CNS Chemistry Department at AstraZeneca Pharmaceuticals LP and held various other positions at AstraZeneca from 1990 to 2006. Dr. Empfield has a Ph.D. in Chemistry from the University of Pennsylvania, a M.S. in Chemistry from Bucknell University and a B.S. in Chemistry from Lebanon Valley College. The Prince Edward Island BioAlliance has named Martin Yuill as its new director, Incubation Services. Yuill is an experienced executive who has led business startups, turnarounds, and restructurings for companies in diverse industries, including: consulting, technology, innovation management, veterinary pharmaceuticals, nutraceuticals, industrial products, pet consumables, and value-added timber products. He has also launched and built several public private partnerships, non-profits, and business incubation programs. Martin most recently served as both president, CEO and director of Incubation and Acceleration at the Greater Peterborough DNA Cluster, and director of Client Services at the Peterborough Region Angel Network (PRAN). In 2008, he launched the ORIC Kelowna Innovation Centre as the first best practice knowledge economy business incubator in the interior of British Columbia, and, in 2013, launched The Cube as the first technology incubation program in eastern Ontario. RepliCel Life Sciences Inc. has promoted Lee Buckler to the position of CEO and president, effective immediately. Buckler has served as RepliCel’s VP of business and corporate development since October 2014. He holds a Bachelors of Education and a law degree from the University of British Columbia. Since 2000, he has been an executive in the cell therapy industry first spending six years in the Stem Cell Technologies group of companies with Malachite Management and then over two years with Progenitor Cell Therapy. In 2008, he started the Cell Therapy Group, a business development consultancy focused exclusively on the cell therapy industry. Buckler is a frequently invited speaker at industry conferences and an oft-quoted commentator on industry events. He has published numerous articles in peer-reviewed

the brain. An expert in the fields of drug metabolism, pharmacokinetics and pharmacodynamics, Dr. Ducharme is also an adjunct professor of pharmacy at the Université de Montréal and sits on several scientific advisory boards.

Lee Buckler journals and trade publications. He is also on the editorial advisory board for the journals, Regenerative Medicine, Cell and Gene Therapy Insights and BioProcess International. Buckler succeeds David Hall, who has retired after five years leading the company. However, Hall will replace Peter Jensen as chairman of the board, while Jensen will remain with the company as a director. Mr. Hall will also continue in an advisory capacity to the company. The board also appointed Lee Buckler as a director, replacing the company’s chief medical officer, Dr. Rolf Hoffmann. This change in directorship enables Dr. Hoffmann to refocus his time on the advancement of RepliCel’s clinical trials and the commercialization of its dermal injector device. Dr. Julie Ducharme has joined Purdue Pharma Canada as its new vice president, R&D. Fluently bilingual, Dr. Ducharme brings to Purdue almost 20 years industry experience in drug discovery and early drug development, including 16 years at AstraZeneca, where she held roles of increasing accountability, culminating as the lead for the company’s R&D strategy for their analgesia portfolio. Most recently, she was general manager of Human Health Therapeutics at the National Research Council of Canada (NRC). Under her leadership, Human Health Therapeutics became a leading portfolio at NRC, working with biotech firms in the development of biologics, vaccines, and delivery of large molecules to

ESSA Pharma Inc. reports that Dr. David Parkinson has stepped in as company president and CEO. He replaces Bob Rieder who announced his departure from the company and resignation from its board of directors. Prior to his appointment, Dr. Parkinson was serving as a director of the company and he will continue acting in that capacity. Dr. Parkinson past roles include: vice president, Global Clinical Oncology for Novartis; vice president, Oncology Development at Amgen; Sr. vice president, Oncology Research and Development at Biogen Idec and as the CEO of the diagnostics company Nodality. Most recently he has been serving as a venture partner at New Enterprise Associates, Inc. (NEA). Prior to joining industry, Dr. Parkinson worked at the National Cancer Institute from 1990 to 1997, serving as chief of the Investigational Drug Branch, then as acting associate director of the Cancer Therapy Evaluation Program. He is a past chairman of the Food & Drug Administration (FDA) Biologics advisory committee, a past member of the FDA’s Science Board, and is a recipient of numerous awards including the FDA’s Cody Medal. BioAmber Inc. announces that George F.J. Gosbee is joining its board of directors replacing Henry “Pete” Linsert, Jr. who resigned his board seat for personal reasons. George Gosbee brings extensive capital markets experience and deep knowledge of the oil and gas industry. Gosbee has been elected a Class 2 director, to hold office until the 2018 annual meeting of stockholders. He will serve on the Nominating and Corporate Governance Committee and the Compensation Committee. His career spans over 20 years in corporate finance, investment banking, global capital markets and advisory. Prior to founding AltaCorp Capital in 2010, he was chairman, president and CEO of Tristone Capital Global Inc., a global energy investment firm which he founded.

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Improve Nucleic Acid Extraction using an Adjustable Tip spacing Pipette

Goal The Thermo Scientific™ KingFisher™ Duo Prime automated purification system offers a fast way to extract high quality nucleic acids in low- to medium-throughput labs. With the Thermo Scientific™ E1-ClipTip™ Adjustable Tip Spacing Equalizer Multichannel Electronic Pipette it is possible to enhance the extraction process by using only one tool to do all of the pipetting - adding samples, reagents and magnetic beads on the deep-well 96-plate or 12-well elution strip can be done entirely using one pipette. The E1-ClipTip Equalizer 6-channel, 15-1250 μL pipette will save

Table 1

valuable lab time and reduce the risk of incurring injuries related to the repetitive motions required in the process.

Introduction The samples for DNA extraction with the KingFisher Duo Prime purification system may vary from plant leaves to mouse ears, or any organism of interest. The sample can be extracted using a variety of sample vessels available; e.g. blood tubes of different sizes, 15 mL/50 mL conical tubes, microcentrifuge tubes or microplate formats. The KingFisher Duo Prime purifi-

E1-ClipTip pipetting function list for dispensing KingFisher Cell and Tissue DNA Kit samples and reagents for KingFisher Duo Prime.

cation system works in standard 24and 96-plate formats and typically there is a need to use several pipettes of different volume settings to fill the reagents on the plate and the elution strip. However, the need to switch between pipettes has been removed. Thermo Scientific KingFisher deepwell 96-plate and KingFisher Duo Prime elution strips can easily be filled using the E1-ClipTip Equalizer pipette. The adjustable tip spacing feature makes it possible to transfer samples from tubes to plates, and dispense magnetic particles and buffers effortlessly with multi-dispensing pipetting functionality. It is also easy to program, save and name a ready-to-use protocol to fill a plate for routine applications in the Programs mode. E1-ClipTip pipettes feature a unique Matrix function that utilizes step-based programming which allows the creation of fully customized pipetting protocols. As such, this provides an ideal option for users that require a variety of pipetting functions in a unique or specific order to complement different research protocols. In this article, the simplicity of using the E1-ClipTip Equalizer 6-channel, 15-1250 μL pipette to dispense samples and reagents on the KingFisher Duo Prime is described with an example of DNA extraction from human cells. One pipette is used for all sample and reagent steps with easy access to useful functions such as mixing and multi-dispensing, to increase pipetting efficiency.

Materials and Methods The E1-ClipTip Equalizer 6-channel,

15-1250 μL pipette was programmed for pipetting all samples and reagents for DNA purification from human cells using the KingFisher Cell and Tissue DNA Kit on the KingFisher Duo Prime purification system. 12 parallel samples of 5 x 105 HeLa cells were used for testing. Cell lysis was performed according to kit instructions and the lysed samples were pipetted from microcentrifuge tubes to the KingFisher deep-well 96-plate using a pre-programmed E1-ClipTip Equalizer 6-channel, 15-1250 μL pipette. Two programs were generated using the Matrix function. One program was created to guide users through the sample row pipetting “KF CT Sample row Duo” and another one for pipetting washing and elution buffers “KF CT Buffers Duo”. Shortcuts can be created for the most commonly used programs on the E1ClipTip main menu for straightforward and quick access in the laboratory. KingFisher Cell and Tissue DNA Kit instructions were followed to adjust steps and volumes on the E1-ClipTip program according to Table 1. Thermo Scientific ClipTip 1250 pipette tips were used in all pipetting steps. The samples and reagents were pipetted on the KingFisher deep-well 96 plate except for the Elution Buffer, which was pipetted on the KingFisher Duo elution strip. In order to achieve uniform results, it is extremely important that a homogenous solution of magnetic beads is reached. To ensure this evenness of bead distribution, the magnetic beads are mixed using the pipette, before aspiration of the solution for multiple

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Figure 1 a

Laboratory Focus February/March 2016

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Agarose gel

Figure 1 b

dispense using a single channel of the E1-ClipTip pipette. The “KF_TissueDNA_Duo” protocol was started on the KingFisher Duo Prime purification system to perform the DNA purification. Once filled, the deep-well 96-plate and 12-well elution strip were inserted into the instrument following the instructions on the user interface. The Thermo Scientific™ Multiskan™ GO microplate spectrophotometer was then used to determine DNA quantity and quality by common absorbance measurements.

Results Genomic DNA (gDNA) purification from HeLa cells resulted in even yields of high quality gDNA. The resulting agarose gel shows clear bands indicating intact gDNA, and the absorbance spectra shows extracted gDNA to be pure without remains of proteins or other impurities (Figures 1a and 1b).

Conclusions The E1-ClipTip Equalizer 6-channel, 15-1250 μL pipette with adjustable tip spacing is an ideal solution for dispensing samples and reagents for nucleic

DNA spectra (A220-A320 nm) of gDNA purified from HeLa cells.

acid purification using the KingFisher Duo Prime purification system. By using the E1-ClipTip Equalizer 6-channel pipette, it is possible to use just one pipette and one size of pipette tip to easily dispense all reagents across all protocol steps, when dispensing volumes from 25 to 800 µL. The adjustable tip spacing helps to reduce repetitive motions, thus increasing personnel safety and lab ergonomics, while reducing pipetting time when working between different labware formats. As a result, laboratory efficiency is increased.

Sini Suomalainen is an Applications Manager with Thermo Fisher Scientific.

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Pharma Notes Axxam SpA (Milan, Italy) and the NEOMED Institute (Montréal, QC) have entered into a strategic drug discovery partnership to identify and develop novel small molecule agonists of FFA4 (also known as GPR120) linked to a range of metabolic and inflammatory diseases. Several pre-clinical studies support the development of a GPR120 agonist as a highly differentiated treatment of diabetes with potential to impact a number of parameters, including protection of the pancreatic β-cells. The collaboration will be based around the optimization of classes of molecules discovered using Axxam’s High Throughput Screening platform and the AXXDIV2.0 compound library. Under the terms of the collaboration, Axxam and NEOMED will integrate their efforts to generate clinical candidates to be developed further to demonstrate clinical proof of

concept. ScarX Therapeutics (Toronto, ON), a Canadian biotech company developing a therapeutic designed to minimize dermal scarring (fibrosis), has closed a $2 million Series A financing. With the proceeds, ScarX says it will complete a Phase 1, single-site clinical trial to establish a safety profile for its lead candidate, SCX001, in human volunteers, with the eventual goal of creating better functional and cosmetic patient outcomes. Twenty-four subjects will participate in the trial, which will take 18 months. SCX-001 is the first nefopam topical cream designed to reduce post-surgical scarring and improve wound healing following surgical wound closure. ScarX Therapeutics is built upon Dr. Benjamin Alman’s 30-year research career, and is based upon

the novel finding that nefopam, a well-characterized small molecule, modulates a protein called beta-catenin that is associated with hypertrophic or excessive scarring in humans. The molecule, originally developed as a prescription oral and IV analgesic in Europe over 30 years ago, had never been applied to dermal scarring until Dr. Alman made a scientific breakthrough by discovering this new application. He invented the technology while at The Hospital for Sick Children (SickKids). The U.S. Food and Drug Administration (FDA) has issued a Complete Response Letter to Telesta Therapeutics (Montréal, QC) Biologics License Application (BLA) for MCNA,stating that an additional Phase 3 clinical trial would be necessary to adequately establish its efficacy and safety. MCNA is a biologic therapy developed to provide high-risk, non-mus-

cle invasive bladder cancer patients who are refractory to or relapsing from first line therapy with bacillus Calmette-Guérin (BCG), with a therapeutic alternative to surgery. It is derived from the cell wall fractionation of a non-pathogenic bacteria. Dr. Michael Berendt, CEO of Telesta, expressed his disappointment with the FDA’s decision, but said the company would continue to work closely with the FDA to assess the costs of a Phase 3 clinical trial. The FDA also encouraged Telesta to meet with them to discuss further clinical development of MCNA. Had it been approved, MCNA would have been the first new bladder cancer therapeutic to reach the market since 1998. AbCellera (Vancouver, BC), a biotechnology company specializing in the rapid discovery of monoclonal antibodies from natural immune cells, has

struck another collaborative deal with big pharma, this time teaming up with Teva Pharmaceutical Industries Ltd. The two sides will collaborate on research using AbCellera’s high-throughput single cell antibody platform to discover rare monoclonal antibodies. Under the terms of the agreement, AbCellera will receive an upfront payment, research payments, and is eligible to receive undisclosed downstream milestones associated with the development and approval of therapeutic antibodies. AbCellera reached a similar deal recently with Merck that would see AbCellera use its high-throughput antibody discovery platform to identify antibodies that specifically modulate target function. That agreement granted Merck the option to develop antibody candidates identified through the collaboration for specified therapeutic applications.

February/March 2016 Laboratory Focus www.laboratoryfocus.ca

FEATURE B Y DANIEL S OM E

Protein Quality Control

in SPR and BLI High-Throughput Screening Studies Summary While surface plasmon resonance and bio-layer interferometry are widely accepted and highly valued tools for screening studies of drug candidate molecules, the data they provide are only as good as the solutions loaded onto the instruments. Preliminary assessment of the quality of the sample proteins and solutions is imperative for reliable binding results. Dynamic light scattering is commonly used to evaluate protein aggregation, degradation and solution quality. However, in the context of high-throughput screening, conventional dynamic light scattering detection is just not feasible, since these instruments work with single-sample microcuvettes and the amount of labour required would be quite extensive. The DynaPro Plate Reader II overcomes this obstacle as it measures dynamic light scattering in situ in industry-standard microwell plates, performing automated, nonperturbative quality assess-ments with minimal time and effort. After analysis on the DynaPro, a ‘heat map’ created by the software offers a quick visual scan of the aggregation state in each well. Detailed particle size distributions may be examined more closely. The operator can readily determine which solutions are suitable for binding assays in order to ensure confidence in the results.

Figure 1. The DynaPro Plate Reader II assesses solution quality in standard 96, 384 or 1536 well plates, without perturbing the samples.

The plates can then simply be transferred to the SPR or BLI instrument with no intermediate fluid handling or perturbation. This seamless work flow greatly enhances productivity in drug discovery.

Introduction Surface Plasmon Resonance (SPR) and Bio Layer Interferometry (BLI) are powerful and widely-used techniques for high-throughput screening and discovery of candidate biotherapeutics. In high-throughput SPR, molecules identified as key targets for treatment are immobilized on a chip, and solutions of potential binding partners injected at one or more concentrations in order to assess affinity and kinetics of interaction. In BLI, the targets or candidates are immobilized on fiber optic probes and dipped into microwells containing solutions of the opposing molecules. The outcome of the screen is the selection of one or more candidate therapeutic molecules with advantageous properties, such as high affinity and rapid binding kinetics. In a typical screen, dozens to hundreds of candidates may be tested, so many high-throughput SPR instruments are designed to draw sample from standard microwell plates and inject into microfluidic channels. The analyte flows over the immobilized target molecule, where good candidates settle rapidly onto the chip surface via specific association with the target epitope. A similar process occurs in BLI, except no microfluidics are involved. An optical probe then provides a signal proportional to the increase in surface-bound mass, and the analysis of these signals over multiple analyte concentrations yields affinity and kinetics. An oft-overlooked obstacle to effective candidate selection is sample quality. The purity and solution prop-erties of molecules employed in SPR and BLI screening may impact the measurements adversely in two ways: 1) data quality and 2) microfluidic integrity. Fortunately, a valuable tool is readily available to address sample quality analysis. High-throughput dynamic light scat-

tering (HT-DLS) with the Wyatt DynaPro® Plate Reader II illuminates protein quality without perturbing the solution, performing measurements in the same microwell plates used by high-throughput discovery screening platforms. HT-DLS enables

rapid evaluation of sample solutions, prior to their loading onto the SPR or BLI instrument, catching material of poor quality before it has the chance to plug the microfluidics or lead to wasting of valuable time and resources on meaningless measurements. As

Figure 2

Sensorgram obtained in the presence of aggregated analyte (simulated).

Figure 3

Aggregates may impact SPR and BLI measurements in several ways.

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Laboratory Focus February/March 2016

FEATURE Figure 4

Figure 5

Analyte self-association with Kd= 100 nM can shift the apparent analyte-ligand equilibrium isotherm from the expected curve (solid line) to the measured curve (dashed line). In this analysis the dimer was assumed to be inactive.

an added benefit, DLS can determine the analyte’s diffusion coefficient, an important property in SPR experiments for identifying mass transfer limitations. More information regarding dynamic light scattering may be found at www.wyatt.com/DLS. The Importance of Analyte Quality I: Impurities Like most techniques, SPR and BLI are subject to the unavoidable, fundamental law of experimental science succinctly put as ‘garbage in, garbage out.’ Analyte quality, so often ignored, is actually quite critical to obtaining accurate and meaningful measurements that ultimately lead to selection of the best therapeutic can-

Figure 6

didates – and hence the best clinical results. Impurities of low molecular weight, such as extractables and leachables, usually have relatively low impact on SPR and BLI measurements. However, large impurities, such as aggregates and foreign particles (both described by the recentlycoined – and very apt – term of ‘nanocrud’, see www.chi-peptalk.com/ biologics-formulation) can wreak havoc on measurements by both techniques. Large impurities contribute to four basic types of experimental uncertainty in SPR and BLI measurements: noisy signals, spurious signals, inaccurate concentra-tions, and skewed kinetics. The evanescent optical fields that

As molecules move through Brownian motion, the light scat-tered from each molecule traverses a different path. This leads to constructive or destructive interference at the light scattering detector. When scattered waves interfere constructively, the DLS detector records high light intensity; conversely when waves interfere destructively, the detector records low light intensity.

probe binding do not extend very far into the solution, typically a few hundreds of nanometers. And yet, any nanoparticle or aggregate passing within that distance from the surface of the chip or fiber probe will result in a signal spike approximately proportional to its mass. Consider a 100 x 100 µm² surface immobilized with bound ligand and illumi-nated by the SPR beam. Exposing this surface to a concentration of analyte >> Kd results in full coverage and a maximum binding signal. Now, consider a single contaminating nanoparticle of ~5 µm diameter in that analyte solution. Since the contaminant contains the same volume as the bound analyte, upon passing very close to the chip

Columns Packed with Fully Porous versus Core-Shell Particles

Autocorrelation analysis is the mathematical transformation linking light intensity fluctuations to the diffusion coefficient. Following autocorrelation, DYNAMICS converts the diffusion coefficients to size.

or probe surface, this particle can generate a noise spike in the binding signal equivalent to all of the bound analyte. Smaller nanoparticles and ag-gregates, that may be present in larger numbers in a sample of low quality, will produce a steady stream of small signal fluctuations leading to a degraded optical response. Both effects are represented in the simulated sensorgrams shown in Figure 2. Analyte aggregates may or may not be active. If active and present in appreciable quantities (e.g., > 5% total analyte protein mass), the aggregates will bind to the immobilized ligand and generate an SPR or BLI signal larger than that of the expected monomeric interaction, skewing the binding response towards a higher estimated affinity and on rate. Aggregates presenting multiple binding sites may exhibit ‘avidity’ effects – interacting simultaneously with multiple immobilized molecules or exhibiting a reduced dissociation rate by hopscotching along the surface of the chip, spuriously leading to an extreme overestimate of affinity. If analyte aggregates are inactive, the effective concentration will be lower than the measured total concentration, leading to decreased binding and an apparent decrease in affinity. Either way, aggregates lead to incorrect quantification of candidate binding properties (Figure 3). The addition of active non-monomeric species with different diffusion properties and binding kinetics than the monomer may also adversely impact the time-dependent sensorgrams. Since the analysis assumes a single binding species with unique on and off rates, the presence of multiple binding species will create binding curves that can-not be fit correctly under the standard assumptions.

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FEATURE Figure 7

Representative size distributions determined by DLS.

The Importance of Analyte Quality II: Self-Associating Analyte Standard SPR and BLI analyses require that the analyte be monomeric in solution at the concentrations employed in the experiment. The previous section addressed the adverse impact of irreversibly aggregated material on the analysis. Poorly formulated or otherwise ‘sticky’ analytes may self-associate reversibly as well as irreversibly. When this is true, the analyte monomer is in dynamic equilibrium with small oligomers, such as dimers or tetramers, and the actual concentration of mono-

aggregated immobilized proteins. In particular, the presence of protein aggregates on the chip or fiber probe surface will most likely lead to a decrease in active material or in the average number of exposed epitopes per immobilized mass. Consequentially aggregated substrate proteins decrease the apparent affinity.

mer varies with protein concentration. Once again the effect on the final measurement depends on the activity and presentation of binding sites, where active oligomers lead to overestimates of affinity and inactive aggregates lead to underestimates (Figure 4). Moreover, presentation of multiple binding sites may lead to avidity effects and gross overestimates of affinity.

Where’s the Drain Opener? Severely aggregated or otherwise impure material bearing large particulates can lead to another highly detrimental effect: clogged microfluidics in multichannel SPR. These fluidic channels tend to be narrow but long and are prone to plugging by agglomerated proteins or other ‘nanocrud.’ The occurrence of clogging events in the middle of a screen of dozens or hundreds of candidates can ruin the efforts of weeks, if not months, of protein expression, purification and preparation – including all the work devoted to analytical method development and assessments of SPR immobilization protocols. Recovery of plugged microfluidics might be as simple as replacing a chip or involve lengthy system cleaning and maintenance. In any case, the damages can amount to many thousands of dollars, even before accounting for lost productivity.

The Importance of Substrate Protein Quality Much as aggregated analytes can lead to experimental errors, so too can

Dynamic Light Scattering to the Rescue! Dynamic Light Scattering (DLS) is a non-invasive, non-perturbative opti-

Figure 8

Visualization of protein quality via heat map in DYNAMICS. Total data acquisition time for 96 wells was < 45 minutes. Data courtesy of Sabin Vaccine Institute and Texas Children’s Hospital Center at Baylor College of Medicine.

cal technique that measures the size distribution of nanoparticles in solution/suspension, from less than 1 nm up to several micrometers. DLS relies on the principles of Brownian motion to determine diffusion rates of particles in solution. The information is transformed by DYNAMICS® software into a particle size distribution which can be evaluated to determine whether or not the solution may safely be injected into SPR microfluidics and whether or not the SPR or BLI measurements will produce reliable results. DYNAMICS provides automated analysis and visualization of DLS results as a heat map indicating good, intermediate, and poor protein quality. The entire process may be completed rapidly prior to loading onto the interaction apparatus simply by transferring the microwell plate into the DynaPro HT-DLS system, running the sample screen, and then (when not contraindicated) loading the same microwell plate onto the SPR or BLI instrument. Microwells that show low-quality material can then be deselected in the interaction screening protocol. Molecules and nanoparticles in solution or suspension ‘jitter’ due to Brownian motion, a consequence of the thermal energy of solvent molecules and the momentum imparted to the nanoparticles by collisions. In DLS, a laser beam impinges on the nanoparticles and is partially scattered in all directions. The light waves scattered by different nanoparticles reach the detectors at different phases and so interfere constructively or destructively at the detector depending on the specific phase difference between them, as shown in Figure 5. The measured intensity of scattered light fluctuates over the time scale characteristic of diffusion. DLS captures the rates of fluctuation to determine transla-tional diffusion coefficients Dt. As described in Figure 6, diffusion coefficients are transformed to particle sizes via the Stokes-Einstein equation: Rh=kBT/6πηDt, where Rh is the particle’s hydrodynamic radius, kB is Boltzmann’s constant, T the absolute temperature, and η the solvent viscosity. Figure 7 shows three typical %-intensity size distribution determined by DLS. The red and blue curves arise from monomodal populations of BSA (Rh = 4 nm) and polysty-rene spheres of radius 50 nm, respectively. The green curve arises from a solution containing a high proportion of monomeric protein, Rh~4 nm, as well as some large aggregates with an Rh of about 50 nm. Because the scattered intensity is proportional to molar mass, the %-intensity curve is heavily weighted toward large particulates, and in fact, the total mass of polystyrene beads is much less than

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Laboratory Focus February/March 2016

FEATURE quality since it will indicate with excellent sensitivity the ‘nanocrud’ content – sub-micron particulates and large protein aggregates that are detrimental to the binding analysis and microfluidic system. The same analysis should also be used to assess dilution buffers employed in SPR to create a series of ligand concentrations.

Figure 9

Conclusions

The DynaPro’s on-board camera helps identify wells containing clean solution and those where sample has precipitated or other-wise can be expected to provide poor data quality.

that of the BSA. The actual amount of aggregate vs. monomeric protein may be estimated via the %-mass size distribution graph (not shown). DLS does not have sufficient resolution to discriminate monomers from dimers or other small oligomers; in general, it can only resolve populations of nanoparticles that differ in size by 3-5x in radius (equivalent to about 100x in mass). However, the presence of small aggregates is inferred via the width of the peak (known as polydispersity) or shifts in the average value of Rh for the popula-tion. HT-DLS Does the Job, Quickly and Easily Traditional DLS takes place in a microcuvette, manually, one sample at a time. It would not be feasible to test all the hundreds of candidates to be screened in this man-ner, though cuvette-based DLS could still be valuable for other quality assays. On the other hand, the DynaPro Plate Reader II brings the power of HT-DLS to bear on quality assessment for the target and all the candidates, thanks to its microwell-plate based format with in situ, non-perturbative measurements. With no fluidics, the DynaPro presents no concern for potential carryover of samples between the wells. Measurements may be completely rapidly, typically requiring 10 to 30 seconds per well including transition time between wells. The entire screen is set up to pro-ceed unattended in the DYNAMICS software package. In HTS-DLS applications, DYNAM-

ICS is usually configured to bin the data as a heat map based on poor, intermediate and high quality size distributions according to bin definitions specified by the user. For example, a sample which shows a single, narrow peak at a size corresponding to that of the analyte may be classified as high quality and allowed to proceed to the binding assay with high confidence (Figure 8, red wells). An adjoining sample which shows a broadened monomeric peak, indicative of some oligomers and perhaps low levels of additional particulates tens of nanometers in size may be classified as intermediate quality and allowed to proceed but with a warning flag as to confidence in the results (Figure 8, blue wells). A sample exhibiting significant particulate content in the micron-size range can be assumed to be either contaminated or highly prone to aggregation and prevented from continuing on to the binding assay (Figure 8, black wells). Additional Benefits As an added bonus, DLS inherently determines diffusion coefficients which are helpful in assessing mass transfer effects: the mass-transfer-limited reaction rate in SPR is km=0.98(D⁄h)2⁄3(f⁄bx)1⁄3, and the diffusion layer thickness is d=D⁄km, where D is the diffusion coefficient, h and b the height and width of the SPR flow cell, respectively, x the distance from the flow cell entrance and f the flow rate.1

Another unique feature of the DynaPro Plate Reader II is the built-in, high-magnification camera which snaps a picture of each well after taking a DLS measurement. These images, stored and shown with the associated DLS data, are especially helpful as diagnostics. A review of the images is useful for determining why the black-classified data in Figure 8 look bad: has the sample precipitated, or perhaps the well inadvertently was not actually loaded with sample? Figure 9 presents addition-al examples of sources of poor data identified by camera images. Finally, the same microwell plates utilized in the DynaPro may be transferred to a spectroscopic plate reader for additional confirmation of content and quality. Sensitivity Yes, every instrument and technique has its limitations on sensitivity. The lower limit of robust detection for the DynaPro is 0.125 mg/mL lysozyme (M = 14.4 kDa). Since the intensity of light scattered by macromolecules is proportional to molar mass, the sensitivity is inversely proportional to molar mass, translating to a lower limit of 0.0125 mg/mL of a 150 kDa IgG. Sensitivity to aggregates follows the same trend, i.e., a 100 nm Rh aggregate consisting of approximately 5000 IgG monomers will be indicated at a concentration of ~ 2 ng/mL. Even if the primary sample concentration is below the limit of detection, DLS is still a useful test of solution

The selection of candidate molecules with the potential for optimal therapeutic effect and patient benefit depends on a reliable target binding screen, as performed with SPR or BLI. This requires, in turn, assuring that the solutions utilized in the analysis are of good quality. Poor quality samples impact the data quality adversely and hold the potential for fouling flow cells and microfluidic channels. High-throughput dynamic light scattering with the DynaPro Plate Reader II is readily implemented in the screening work flow to classifying solutions as 1) high quality, offering maximal confidence in the interaction analysis; 2) intermediate quality, suitable for measurement with caution in relying on the results; and 3) low quality, not suitable for analysis and potentially fouling the measurement device. Adding an HTDLS pre-screen can prevent much of the uncertainty and productivity loss associated with variable ligand quality, leading to more reliable binding data and confidence on the final candidate selection. After identification of the most promising candidates, the DynaPro is also widely used in pre-formulation and candidate developability studies to assess aggregation, conformational and colloidal stability.2,3 See www.wyatt.com/DLS for additional information.

Daniel Some, PhD. is a Principal Scientist with Wyatt Technology Corp.

References 1. Karlsson et al. Methods 1994, 6, 99-110. 2. Saito et al. Pharmaceutical Research 2013, 30(5), 1263-1280. 3. Razinkov et al. Current Drug Discovery Technologies 2013, 10(1), 59-70.

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feature

b y John D Ma cFarl ane

Protein Separation in reversed-phase mode Chromatographic separations of proteins are based on their difference in either size (size exclusion chromatography), electric charges (ion exchange chromatography), or hydrophobicity (reversed-phase chromatography).

Shiseido’s “Proteonavi column” was developed to separate proteins and peptides in reversed phase mode. In reversed-phase mode, it is know that proteins and peptides with higher-order structure are denatured to some extent in course of being retained on the stationary phase after introduction to the column. Once the retention with denaturation has occurred, molecules will not migrate until the organic content of the mobile phase is raised to an appropriate level under a gradient program. In reversed-phase separation, peak shapes are often deteriorated when secondary interactions, such as electrostatic interactions with partial bare silica or metal imprities, exist between the stationary phase and proteins. Listed below are common facts in analytical and preparative separations of proteins and peptides. 1. Stationary phase with long alkyl chains (C18) shows irreversible adsorption of protein. 2. Stationary phase with shorter alkyl chains (C1, C4, or C8) have a lower durability under acidic mobile phases. 3. Stationary phase with shorter alkyl chains are influenced by silica, as a starting material, showing a lot of variation. 4. Stationary phase with shorter alkyl chains often shows insufficient separation.

All of the above issues are related to chemistry and to introducing alkyl chains to silica. The type of alkyl chain and the way to anchor it on silica seem to be the keys of preparing an appropriate stationary phase for protein and peptide analysis. Proteonavi has a C4-modified surface designed after comprehensive basic research.

2. Properties of Proteonavi 2-1 Property values Proteonavi utilizes high-purity silica with few metal impurities and shows minimal irreversible adsorption for proteins and peptides. Its pore size is as wide as 30nm, meaning large pro-

Figure 1

teins are able to have enough interactions with the stationary phase. (See Table 1) 2-2 Evaluation of hydrophobicity and surface polarity The nature of reversed-phase columns can be studied by measuring separation factors of different pairs of standard compounds. The strength of hydrophobicity can be expressed as a separation factor of toluene and benzene, showing a retention increase generated by one methyl group. Similarly, surface polarity is calculated between methyl benzoate and benzene, observing an influence of ester moiety on retention.

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Table 1

Particle size (µm)

Figure 3

feature

Property values

Functional Pore size group (nm)

Figure 2

Laboratory Focus February/March 2016

Specific surface area (m2/g) 105

Carbon content (%) 3.3

Alkyl pH group density (µmol/m2) 3.2 2-10

USP

L26

Results obtained with CAPCELL PAK SG 300 C1, C8, and C18, Proteonavi and two other commercially available columns for protein separation were plotted on the hydrophobicity/ surface polarity graph in Figure 1. Hydrophobicity is the most fundamental parameter in reversedphase columns. Generally, a stationary phase with large carbon content (larger alkyl density and/or longer alkyl chain) shows a large value. A large hydrophobicity is preferred when a large retention or high loadability is required. A large value, however, may not be always good for macromolecules with a structure of higher degrees, such as proteins, since a long alkyl chain (C18) is more likely to cause an irreversible adsorption accompanied by their denaturation. Surface polarity contributes to the retention and separation of polar compounds, and therefore, generates selectivity. Residual silanol groups on silica surface are thought to be one of the sources of surface polarity. While peak shapes of neutral compounds are hardly influenced by them, acidic silanol groups, if any, are known to cause a peak tailing for those having a basic molety, such as proteins. CAPCELL PAK SG300 columns form a characteristic curve in the graph, showing the influences of alkyl chain length on both values. However, even C1, the one having the shortest chain, shows smaller polarity values than those of other product (V and Y), which can be attributed to the poly-

mer-coated surface of CAPCELL PAK series. Proteonavi shows the surface polarity close to that of CAPCEL PAK C1, in spite of its C4 structure. Its entirely different surface coating process provides a new balance of hydrophobicity and surface polarity for protein and peptide analysis.

3. Features of Proteonavi 3-1 Specifically large retention of proteins Proteonavi, having C4 chains, showed overall retention of standard neutral small compounds, which is somewhat between C1 and C8 columns, as expected. However, it retains six standard proteins more than the C8 column, apparently because of its structure designed specifically for protein separation. The large retention does not contradict with the unique property values discussed at the section before. (See Figure 2) 3-2 Outstanding acidic durability Acidic mobile phases containing trifluoroacetic acid (TFA) are often used in protein separation in reversedphase chromatography. Acidic hydrolysis of alkyl chain is known to occur more or less on any silica-based columns, which is more pronounced for those with shorter alkyl chains. Partial loss of alkyl groups generally causes a decrease of overall retention of analytes and a change of selectivity toward them. In order to evaluate durability of different columns under acidic conditions, an accelerated test was performed in the conditions show below. Retention time change of benzylalcohol was examined under the strongly acidic mobile phase at the elevated temperature. As shown in Figure 3, all of the columns tested showed loss of reten-

Figure 4 Actual chromatograms of the first and 20th runs (10th run for column Y) are shown below. The absence of change in uracil peak (shape and position) at to indicates that the test hardly damaged silica support in all of the columns. Retention time of benzyl alchohl, on the contrary, showed a significant difference from one column to another.

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Feature Figure 5

Figure 7

kyl chains in the stationary phase. When secondary interactions, such as coulombic interactions by acidic silanols or metal impurities in the stationary phase, exist to a large extent, peak shapes of proteins will be deteriorated due to their slow kinetics. Proteonavi shows better peak profiles and higher resolution among standard proteins, in comparison with conventional widepore columns. (See Figure 5)

Figure 6

3-4 Excellent lot reproducibility Lot-to-lot variation in separation is often discussed in protein analysis. Proteonavi has excellent reproducibility of retention time and separation. (See Figure 6)

tion as time passed. While column Y showed a rapid drop and V lost approximately 30 per cent of its retention during the process, Proteonavi kept more than 90 per cent of the initial retention time until the end.

3-3 Minimal protein adsorption and high column efficiency The primary source of protein retention on reversed phase is hydrophobic interaction between hydrophobic parts of amino acid residues and al-

3-5 Minimal column bleed “Column bleed” used to be a term used in gas chromatography with universal detectors, such as flame ionization detector. The recent development of interfacing technologies in mass spectrometry brought the term into the field of Liquid Chromatography (LC). Column bleed of an LC column is a synonym of a signal level of total ion chromatogram (TIC) obtained with LC-MS under gradient conditions at full scan mode. The source of column bleed is a bonded phase on silica. It may affect ionization efficiency of analytes of interest, or will be potential impurities in preparative separation. Proteonavi was designed to minimize

the level of column bleed. A comparison among conventional wide-pore columns and Proteonavi is shown below. These background chromatograms were obtained under gradient conditions without sample introduction. (See Figure 7)

Conclusion Shiseido’s Proteonavi columns have a C4-modified surface that has been designed based on a comprehensive research program. Thus, the Proteonavi series of columns have been shown to be very durable in acidic mobile phases despite the use of a short alkyl chain. Excellent lot reproducibility combined with minimal protein adsorption, high column efficiency and minimal column bleed makes these columns ideal for all protein and peptide separations. Proteonavi columns are also suitable for scaling up from analytical to preparative separations with column inner diameters up to 30 mm available.

John D MacFarlane is General Manager, at JM Science Inc.

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Clamps BioPure Technology, part of Watson-Marlow Fluid Technology Group, announces the release of its BioClamp® patented plastic union tri-clamp. The clamp is designed to meet the specific needs of pharmaceutical and bioprocessing laboratories. The clamp is able reduce distortion on polymeric fittings when subjected to heat and a tamper-evident feature is available on the 1/2” to 8” products, delivering full confidence in the clamp’s integrity. Moreover, the BioClamp is molded from reinforced Nylon 66 USP Class VI, FDA making it suitable for sterilization by autoclave and gamma irradiation. The clamp is also manufactured and packed in an ISO Class 7 clean room and has lot numbers molded in for full traceability.

Web: www.watson-marlow.com

Pipettes Sartorius has introduced a new range of mechanical pipettes called Tacta. The new pipettes are available in a range of volumes from 0.1 to 10,000 μl in single channel models, and from 0.5 to 300 μl in multi-channel models. Its new Sartorius Optiject, levered tip ejection technology, enables controlled and smooth tip ejection with minimum force. The Optiload feature, with spring-loaded tip cones ensures tip loading with perfect sealing. In addition, the new Sartorius Optilock system provides flexible volume adjustment and locking functionalities, and prevents accidental volume changes during pipetting. An added feature includes a clear 4-digit display, which makes the volume easy to read, even when the pipette is at an angle. Moreover, its integrated adjustment functionality and scale show the degree of adjustment and, by noting this value for a specific liquid, the user can return to that setting any time. With only three parts to disassemble and requiring no tools, Tacta pipettes are also particularly quick and easy to clean.

Web: www.sartorius.com

Chromatography The Thermo Scientific Dionex Integrion High-Pressure Ion Chromatography (HPIC) system breaks new ground in productivity and efficiency for environmental, food safety, pharmaceutical and industrial/petrochemical laboratories. The new system delivers features previously available only on Thermo Scientific high-end systems, including high-pressure capability and optional electrochemical detection. With a simple, logical, flow-based plumbing layout and integrated performance features, including whole-system smart monitoring, the Dionex Integrion HPIC offers fast run times. Other system features include easy-to-install IC PEEK Viper Fittings that minimize peak dispersion and band broadening—ultimately improving chromatographic resolution, a detachable tablet with local language support that allows the flexibility to access IC controls even while away from the instrument and finally a thermally regulated detector compartment that provides extended life to consumables.

Web: www.thermoscientific.com

New Products Benchtop Analyzer The Altair™ 240 from EKF Diagnostics is a compact, reliable and fully automated random access benchtop analyzer. With positions for 43 reagents and the ability to configure open channels for your esoteric testing needs, it provides a comprehensive solution tailored to meet the needs of your laboratory. The device is supported by a full-range of bar-coded, liquid-stable, ready-to-use Stanbio Chemistry reagents. The Altair™ 240 automatically reads and manages barcoded reagent bottles and sample tubes. Most reagents have 30-day on-board stability to minimize waste. The intuitive Windows®-based software screens make learning and operating the analyzer quick and easy. Additionally, the device can run up to 400 tests per hour with an optional Ion Sensitive Electrode (ISE) module. Dual reagent probes translate to higher throughput and faster patient turn-around time.

Web: www.ekfdiagnostics.com

Scanners Ziath, maker of sample tracking solutions for use in laboratories and biobanks, has released the HandHeld as the newest addition to its line of scanners. The intelligent device is portable and provides users with the ability to immediately identify tubes and track samples when away from the lab. This includes manual picking from repositories, as well as effective recording when collecting samples in the field. The scanner can be operated using one hand and incorporates a lightweight build and intuitive interface that’s easy enough for anyone in the lab to use. Its portable nature is perfect for a number of underserved applications, including the generation of multiple pick lists for accurate tube selection, vial checking to confirm your sample is what it says it is, and the ability to easily record samples in the field at the point of collection. Since the scanner is battery operated, users have the option to effortlessly enter data about specific samples as soon as they’re scanned – a feature that generally requires the scanner to be connected to a central computer system.

Web: www.ziath.com

Sample Preparation NuGEN Technologies Inc. announces the launch of its Ovation® SoLo RNA-Seq System, the company’s latest sample preparation tool. The Ovation® SoLo kit facilitates research in areas such as blood-borne biomarkers, cellular heterogeneity, and the study of single cell transcriptomics, while providing a complete end-to-end solution for whole transcriptome analyses starting directly from cells or isolated RNA. The system features an improved method for ribosomal depletion of > 90%, efficient cDNA generation from ultra-low inputs that provides a complete representation of the entire transcriptome, and the capability to unambiguously identify PCR duplicates.

Web: www.nugen.com

New Products

February/March 2016 Laboratory Focus www.laboratoryfocus.ca (488nm) and red (640nm) lasers, along with 15 detection channels. The expanded range of dyes and fluorochromes for the violet laser also adds flexibility and enhanced performance, with lower spectral overlap, increased brightness, better signal-to-noise ratio and broader applicability.

Sample Preparation IntelliCyt introduces the iQue Screener PLUS Screener platform, an integrated instrument, software and reagent system that enhances the screening workflow, from sample preparation to results. It enables rapid, high-content, multiplexed analysis of cells and beads in suspension in 96-, 384- and 1536-well plates. The system’s patented sample delivery system enables rapid plate processing (less than five minutes for 96 wells), and assays can be miniaturized to conserve precious sample and reduce reagent use. Softwareassisted automation, analysis and experiment-level visualization tools reveal deep insight into complex biology through an easy-to-use, intuitive interface. The iQue Screener PLUS also expands choice and flexibility, adding a 405nm violet laser to the blue

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Web: www.intellicyt.com

Chilling/Heating dry baths Torrey Pines Scientific, Inc. announces the release of its EchoTherm™ Models IC30 and IC30XT, Peltier-driven high capacity chilling/ heating dry baths capable of handling a large variety of sample blocks and sample capacities available. The IC30 (-10°C to 100°C) and IC30XT(-20°C to 100°C) can freeze, chill, or heat samples in a variety of sample blocks that can hold 0.2 ml to 50 ml centrifuge tubes, test tubes, vials, assay plates, and even round-bottom flasks. The units can freeze, chill, or heat 64 to 1.5ml centrifuge tubes, 9 to 50ml centrifuge tubes, or even 4 to 250ml flasks. These are just some of the large variety of sample blocks avail-

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able. The units have digital display and control to 1°C, 30-day count down timer in hours/minutes/seconds, data logger, and RS232 I/O port to collect data or to control the units by computer. The IC30 and IC30XT measure 8.5” (216 mm) wide x 10” (245 mm) deep x 4” (102 mm) tall. They come complete with chiller/heater module, universal power supply, AC line cord for the country of use, and the instruction manual. They are UL, CSA, and CE compliant.

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Columns The new Thermo Scientific mass spectrometric immunoassay (MSIA) Strepdavidin EVO microcolumns are designed to help scientists analyze large biomolecules, such as proteins and antibodies, with improved accuracy, consistency and simplicity, even at high throughputs. When combined with the Tecan Freedom EVO robotic platform, MSIA Streptavidin EVO microcolumns enable fast and accurate isolation of target analytes from complex biological matrices. Housed within a pipette tip, the proprietary monolithic columns are densely coated with streptavidin for analytical affinity purification of any biotinylated affinity ligand, even at low concentration. Since this technology is incorporated into a pipette tip, users can analyze sample volumes as low as 10 µL, saving precious samples. Furthermore, when compared to traditional resin and bead-based methods, MSIA technology provides lower background noise and improved analytical sensitivity, while the simple workflow enables easier method standardization and transfer between labs.

Web: www.thermoscientific.com

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Benchtop system Sartorius Stedim Biotech announces the launch of its SARTOFLOW® Smart benchtop crossflow system for optimized ultra and diafiltration applications. Ideal for use in many downstream processes, such as purification of vaccines, monoclonal antibodies and recombinant proteins, the system is suitable in laboratory environments for process development and clinical trials as well as for cGMP environments. It comes equipped with a low shear 4-piston membrane pump that enables high product yields. In addition, the pump provides a wide range of flow rates allowing to choose between mem-

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products from a variety of biological particulates, serving as the primary barrier in life science research and experimentation. As a Class II Type A2 Biosafety Cabinet, the Purair BIO minimizes and contains Biosafety Level 1-3 agents, maintaining negative pressure inside the cabinet during operation to prevent contaminants from escaping the work area. HEPA filtration scours 70% of the incoming room air to protect the products, while the remaining 30% of the exhausted air is filtered by a second HEPA filter. The cabinets provide ample workspace with environmentally sound operation and low energy consumption.

Web: www.airscience.com

SAVE TIME brane surface areas from 50 cm² to as much as 0.14 m². Its touchscreen offers instant access to all critical process parameters including display control and alarm functions. A logbook function stores alarms, set points and user logs. Users can select predefined parameters to automatically run sequences for concentration, diafiltration, rinsing, filling, draining, flushing steps and tare functions. An optional peristaltic diafiltration pump is available to load product or buffer as a discrete process step. The system can also be upgraded in the case of changing process requirements.

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Web: www.sartorius.com

Conjugation kits Innova Biosciences has released new red, black and blue 400nm LATEX conjugation kits for use in the diagnostics field and across other applications. The new conjugation kits are easy to use, and offer quick, effective conjugation of antibodies, proteins and peptides to latex beads, offering the potential to simplify and improve assay development whilst reducing development timelines. Additionally, as a variety of colours are available, the range enables multiplexing and therefore maximum flexibility in assay design. The kits are suitable in lateral flow assays, and across a range of other applications, and are fully compatible with human serum.

Web: www.innovabiosciences.com

Safety Cabinets Air Science has introduced its new line of biological safety cabinets. The Purair BIO is designed to protect individuals, the environment, and

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Laboratory Focus February/March 2016 www.laboratoryfocus.ca

app review PopRX

By: PopRX https://www.poprx.ca/

Promise of a Zika vaccine The global race to take down the Zika virus is beginning to heat up and Canada’s top infectious disease facility is leading the charge. To date, the mosquito-borne virus has reached pandemic status, spreading rapidly in South and Central America. It has also been speculatively linked to birth defects, known as microcephaly, in Brazil. Currently, there are no treatments or cure for the disease, however, the Winnipeg-based National Microbiology Laboratory (NML) believes it will have an experimental vaccine ready by year’s end. While that may sound like a lofty goal to some, for those who know NML’s history, it is by no means an empty promise on their part. Since it first opened in 1999 as Canada’s only facility with a biosafety level 4 laboratory, the NML has been at the cutting edge of both infectious disease research and vaccine development. For starters, in 2014, it played a key role in the global effort to stop the deadly Ebola outbreak in West Africa. In fact, its scientists helped design both the first Ebola vaccine and the Ebola drug ZMapp. Additionally, in 2009, the NML was the first to confirm H1N1, or “swine flu” as a new strain of influenza. And prior to that, in the early 2000s, it helped put an end to the deadly SARS epidemic. The success of the NML goes beyond just its bricks and mortar. It is home to excellent scientists, who produce world class work. Moreover NML scientists collaborate with some of the best infectious disease researchers in the world. Such is the case in the development of a Zika virus vaccine, where NML has partnered with researchers in the United States. In fact, the vaccine itself is already in clinical production. On the Canadian front, the NML is also working with a team of scientists at Brock University in St. Catherines, ON, the only academic institution in Canada with a Level 3 containment lab that includes an insectary. There they have started testing mosquitoes native to Canada to determine whether the insects can become infected with Zika and potentially transmit the virus to humans. Likewise, the national lab is also testing Zika in about a dozen mosquito species, about half of them unique to Western Canada. In both instances they hope to answer questions as to whether the virus could come to Canada. If it does, expect the NML to be on the front lines protecting Canadians.

PopRx, dubbed the ‘Uber’ of prescriptions, is a free app that delivers your prescription medications with just a picture on your phone. It is the first and only Canadian company that offers this service, allowing customers the freedom to get their prescriptions, refills and pill reminders for free with a simple click on their phone. Whether you are a busy parent, or have ongoing prescription needs, you no longer have go to the pharmacy and wait in lines for your medications. PopRx delivers right to your door.

Science Mobile

By American Association for the Advancement of Science http://content.aaas.org/mobile/ The Science Mobile app is a great information source, but falls short in terms of user experience. Starting with the good, the app lets users read summaries, abstracts, and, for subscribers, full-text articles from Science, Science Translational Medicine, and Science Signaling websites. Users can also share their own articles, links, and job postings to the app through Facebook, Twitter, and e-mail. This makes it an ideal information source for chemistry, environmental, clinical and biological science researchers and also a useful tool for job seekers who can easily sift through job listings as well as tap into other skill building resources via Science Careers. As an added bonus, the App gives users access to the Science weekly podcast and other multimedia. The drawbacks of the App are that users have found loading times and app functionality cumbersome and confusing. Many users comment that page loading speeds are terrible, and that the App hasn’t been updated in a very long time. There’s also a lack of tech support to help address troubleshooting problems.

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Laboratory Focus February/March 2016

MARCH 2016 March 6-11 PITTCON 2016 Venue: Atlanta, GA Tel: (412) 825-3220 Fax: (412) 825-3224 email: info@pittcon.org Twitter: @Pittcon Web: www.pittcon.org

March 15 – 16 Bio Asia Venue: Tokyo, Japan Web: www.bio.org/events/ conferences/about-bio-asia

Calendar

May 11-12 Agri Investment Forum Venue: Toronto, ON Tel: (587) 350-0067 Email: matthew@ criticalpathgroup.com Web: www.agri-investmentforum. com

JUNE 2016 June 5-9

and Exhibition Venue: Halifax, NS Tel: 613-232-6252 Web: www.csc2016.ca

June 6-9 2016 BIO International Convention Venue: San Francisco, CA Web: www.convention.bio.org/2016

Canadian Chemistry Conference

JULY 2016 July 17 -21 International Conference of Biochemistry & Molecular Biology Venue: Vancouver, BC Tel: 604 681 2153 x 109 Fax: 604 681 1049 Email: IUBMB2016@icsevents.com Website: iubmb2016.org

March 29-30 World Vaccine Congress 2016 Venue: Washington, DC Tel: +44 20 7827 5945 Web: www.terrapinn.com/ conference/world-vaccine-congresswashington/index.stm

March 30-April 2 AMMI Canada - CACMID Annual Conference 2016 Venue: Vancouver, BC Tel: 613-260-3233 Fax: 613-260-3235 Email: info@ammi.ca Web: www.ammi.ca

Give our kids every chance to get better.

PUT YOUR MONEY WHERE THE MIRACLES ARE.

APRIL 2016 April 4-6 BIO Europe Spring 2016 Venue: Stockholm, Sweden Web: www.ebdgroup.com/bes/ index.php

JENNIFER LOPEZ ACTRESS, MUSICIAN, TV PERSONALITY, MOM

April 17-20 World Congress on Industrial Biotechnology Venue: San Diego, CA Web: www.bio.org/events/ conferences/world-congressindustrial-biotechnology

April 21 18th Annual LifeSciences BC Awards Venue: Vancouver, BC Tel: 604.669-9909 ext. 104 Web: www.2016lsbcawards. eventbrite.ca

April 25-27 MEDEC 2016 MedTech Conference Venue: Toronto, ON Tel: 416-641-2747 Email: dgates@medec.org Web: www.medec.org/events/EventDetails.aspx?id=691028&group=

MAY 2016 May 2-3 Bloom Burton & Co. Healthcare Investor Conference Venue: Toronto, ON Tel: 416-640-7580 Email: bbloom@bloomburton.com Web: www.bloomburton.com/ conference/

Like all moms, I’m always concerned about my children’s well-being. But sometimes they get sick. Sometimes they get hurt. That’s why I’m so grateful we have children’s hospitals. If any child needs a miracle, they’ll do everything in their power to make one happen. Please join me in giving sick and injured children every chance to get better. Put Your Money Where the Miracles Are. Give to your Children’s Miracle Network member hospital. Children’s Miracle Network® raises funds and awareness for 170 member hospitals, 14 of which are in Canada. Donations stay local to fund critical treatments and healthcare services, pediatric medical equipment and research. Its various fundraising partners and programs support the nonproﬁt’s mission to save and improve the lives of as many children as possible. Find out why children’s hospitals need community support, identify your member hospital and learn how you can Put Your Money Where the Miracles Are, at childrensmiraclenetwork.ca and facebook.com/CMNHospitals.

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12/17/15 1:59 PM

NEW: CryoCube® ULT Freezers

Efficiency Reinvented CryoCube Ultra-low Temperature Freezers CryoCube ultra-low temperature freezers combine maximum sample security with improved functionality. New advancements decrease power consumption and make CryoCube freezers among the most energy efficient in the industry. Eppendorf quality means years of trouble-free operation and dependable support.

> New automatic vent port on front door allows for easy re-entry into your freezer while also improving energy efficiency > New ergonomic handle requires less force and improves freezer access > New magnetic closures on insulated inner doors further increase ease-of-use

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Full page tabloid ad Laboratory Focus – LFO

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Laboratory Focus February/March 2016

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