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UBC researcher creates ‘living’ solar cells to convert light into energy
Overcast skies have posed problems for gathering solar energy in places like British Columbia that frequently have clouds on the radar, so researchers from the University of British Columbia took a stab at making a cheap and sustainable way to build solar cells using bacteria to convert light into energy. These cells generated a current stronger than any previously recorded from such a device and worked just as efficiently in dim lighting. With further development, these solar cells—called “biogenic”
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because they are made of living organisms—could become as efficient as the synthetic cells used in conventional solar panels. “Our solution to a uniquely B.C. problem is a significant step toward making solar energy more economical,” says Vikramaditya Yadav, a professor in UBC’s department of chemical and biological engineering who led the project. Solar cells are the building blocks of solar panels. They do the work of converting light into electrical current. Previous efforts to build
biogenic solar cells have focused on extracting the natural dye that bacteria use for photosynthesis. It’s a costly and complex process that involves toxic solvents and can cause the dye to degrade. Yadav’s solution was to leave the dye in the bacteria. They genetically engineered E. coli to produce large amounts of lycopene—a dye that
gives tomatoes their red-orange colour and is particularly effective at harvesting light for conversion to energy. The researchers coated the bacteria with a mineral that could act as a semiconductor, and applied the mixture to a glass surface. With the coated glass acting as an anode at one end of their cell, they generated a current density of 0.686 milliamps per square centimetre—an improvement on the 0.362 achieved by others in the field. “We recorded the highest current density for a biogenic solar cell,” says Yadav. “These hybrid materials that we are developing can be manufactured economically and sustainably, and with sufficient optimization, could perform at comparable efficiencies as conventional solar cells.” The expenditure and cost savings are difficult to estimate according to Yadav, but he predicts the process will reduce the cost of dye production to one-tenth compared to the usual. To see this story online visit https://laboratoryfocus.ca/ubcresearcher-creates-living-solarcells-to-convert-light-into-energy/
PlantForm collaborates to manufacture biosimilar to the South African market Guelph-based PlantForm Corporation of Canada is stretching across the ocean to collaborate with South Africa’s Council for Scientific and Industrial Research (CSIR) and BGM Pharmaceuticals to produce a biosimilar of trastuzumab using PlantForm’s vivoXPRESS® manufacturing system.
This agreement will permit the CSIR to produce a biosimilar monoclonal antibody (trastuzumab) using the tobacco plant-based vivoXPRESS® expression technology on a laboratory scale. The CSIR will demonstrate the utility of PlantForm’s proprietary technology to Continued on page 3
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August/September 2018 Laboratory Focus www.laboratoryfocus.ca
news Canadian Vaccine Catalogue launches to digitize vaccination records As the world moves forward with digitizing data, a team of Canadian physicians, researchers, and policy-makers release the first version of the Cana-
dian Vaccine Catalogue (CVC) to access vaccination records. This comprehensive, standards-based source-of-truth is a crucial resource for vac-
cine terminology and vaccine product information in Canada. It was developed by The Ottawa Hospital mHealth Lab and is funded by the Pub-
lic Health Agency of Canada. Provinces and territories across Canada are working towards the development of a national network of immunization registries whereby each jurisdiction would maintain its own system for tracking immunization coverage. To facilitate Continued on page 3
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news PlantForm Continued from page 1 produce a trastuzumab drug candidate with the long-term goal of manufacturing trastuzumab, and other possible drug candidates, in South Africa for sale in African markets. “This agreement represents a significant opportunity for PlantForm,” says Dr. Don Stewart, PlantForm’s president and CEO. “The South African pharmaceutical sector is the largest in the region, and by working with the CSIR and BGM Pharmaceuticals we continue to advance our global strategy for biosimilar drug development that will make life-saving medications more widely available at much lower cost.” The vivoXPRESS® system is revolutionary, using fast-growing tobacco plants to produce a wide range of biologic (large molecule) drugs, including monoclonal antibodies and therapeutic proteins, at a fraction of the cost of manufac-
Vaccine Catalogue Continued from page 2 the standardization of these registries and other digital health records that collect immunization data, like electronic medical records, the CVC provides monthly updates of vaccine terminology and product information that is aggregated from various governmental and industry sources. The CVC connects a physician’s electronic medical record with an up-to-date list of vaccines that are available for use in Canada and will enable the software to produce vaccination records that are consistent with nationally agreed upon data standards. The CVC also facilitates vaccine barcode scanning for both inventory management and immunization administration. “Our national immunization strategy depends on our ability to collect highquality data on immunizations, in all provinces and territories, and from a myriad of digital systems and providers. The Canadian Vaccine Catalogue can help facilitate this,” says Dr. Kumanan Wilson, senior scientist at The Ottawa Hospital, a professor at the University of Ottawa and founder of The Ottawa Hospital mHealth Lab, based at Algonquin College. This catalogue will become a great tool for all Canadians to keep track of and be aware of available vaccinations in their province or territory. To see this story online visit https://laboratoryfocus.ca/canadian-vaccine-catalogue-launches-todigitize-vaccination-records/
turing the same drugs using conventional methods. Studies have shown that PlantForm’s plant-produced trastuzumab drug candidate is equally as effective at inhibiting the growth of HER2-positive breast cancer cells as the commercial brand-name antibody Herceptin. “We’re pleased to be working with PlantForm and BGM Pharmaceuticals to further development of the vivoXPRESS® technology,” says Dr. Tsepo Tsekoa, who heads the CSIR’s biomanufacturing technology demonstration research group. “The CSIR is committed to utilizing and enhancing our capability in biologics production to assist South African industry with development and localization of cost-effective technology solutions for biopharmaceutical manufacture. This collaboration will strengthen this commitment and help us expand our research into using plant-based processes to develop biologics.” Looking farther into the future, the collaboration will advance BGM Pharmaceuticals’ mission to develop
new biopharmaceuticals and improve existing therapies for emerging African markets, while at the same time showcasing some of Canada’s innovative and forward-thinking technologies. “We’re very excited about the opportunity to bring PlantForm’s lowcost manufacturing platform to the South African market to help make life-saving biopharmaceuticals more
affordable and more widely available throughout the region,” says Martin Magwaza, BGM’s president and CEO. To see this story online visit https://laboratoryfocus.ca/ plantform-collaborates-to-manufacture-biosimilar-to-the-southafrican-market/
Urotronic’s drug-coated balloon catheter receives license for urethral stricture treatments in Canada
Plymouth-based Urotronic receives license to begin treating urethral strictures with its innovative Optilume drug-coated balloon (DCB) in Canada. The development is expected to have a significant impact across Canada where just a handful of physicians currently perform the majority of all stricture treatments. “We are quite excited to see this come to market,” says Dr. Conrad Maciejewski, a reconstructive urologist who treats patients at The Ottawa Hospital and is one of just 9 doctors in Canada who practice in the field of reconstructive urology. Millions of men around the world suffer from incapacitating urethral strictures. The condition blocks the pathway for urine to exit the body from the bladder and can result in a painful, frustrating slowing of the urinary system. Strictures can be caused by infections, trauma and other medical procedures that injure the lining of the urethra. The number of urological procedures is growing rapidly due to a growing elderly
population that are more susceptible to urinary tract problems. Urortonic’s innovative device called Optilume combines balloon dilation with the delivery of an anti-proliferative drug to prevent recurrence of the blockage. In clinical trials performed in Latin America, the drug-coated balloon has performed as intended in both opening blockages and preventing the formation of scar tissue which can develop quickly after any medical intervention. Dr. Maciejewski says there is a significant need for new and alternative urethral stricture treatments. Most of his patients would prefer to avoid major reconstructive surgery called urethroplasty and are also not satisfied with the 40 per cent success rate of a common endoscopic intervention known as an urethrotomy. “This is the first time we’ve had a product that can significantly exceed those success rates with better durability and potentially less side effects compared to a major surgery,” he says. “We are excited to introduce our
innovative drug coated balloon to the Canadian market and look forward to working with physicians and their patients who suffer from this debilitating disease.” says David Perry, president and CEO of Urotronic. Urologists across Canada, many whom will be attending the upcoming Canadian Urological Association (CUA) conference in Halifax, Nova Scotia, are also encouraged by the potential of Optilume™. The device has the potential to lift the burdens of the current Canadian market by adopting a treatment that is easy to learn and can be performed in a short outpatient procedure. “If we were able to offer, in the hands of general urologists or even specialists, something less invasive, more accessible and with better results than what our current less invasive options provide, that would be a great option and, I think, a lot of people would seriously consider it,” Dr. Maciejewski says. A Canadian post-market trial is currently scheduled. The U.S. Food and Drug Administration also continues to monitor clinical trials involving the Optilume DCB in the United States. To see this story online visit https://laboratoryfocus.ca/urotronics-drug-coated-balloon-catheter-receives-license-for-urethral-stricturetreatments-in-canada/
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news
Mega-corporations to propel the life sciences industry towards $1.5 trillion milestone Mega-corporations like Google, Apple, Facebook, and Amazon have not only taken the world by storm but are propelling the life sciences industry towards hitting the $1.5 trillion milestone by 2022. Health care digitization and consumerization, a tsunami of patient data and the emergence of value-based reimbursement models are shaking things up in this ever-evolving industry. The convergence of biopharmaceuticals, drug delivery devices, and companion diagnostics is driving regulatory and commercial changes that make this a very exciting time, according to Frost & Sullivan’s recent analysis, Global Life Sciences Industry Outlook, 2018. “The scope for synergies among disciplines is attracting big tech companies such as Google, Amazon, Facebook, Apple (GAFA) to the life sciences industry,” says Unmesh Lal, program manager, Transformational Health. “The adoption of artificial intelligence (AI) and cloud-based solutions for drug discovery and clinical trial workflow is improving the overall efficiency of production. Additionally, pathology and research laboratories are increasingly adopting health analytics solutions to track the test utilization
• Liquid biopsy and companion diagnostics: Roughly $500 million in venture capital funding is expected to flood the liquid biopsy space as companion diagnostic-based liquid biopsy tests gain approval from the regulatory authority; and • Biotech expansion into CAR-T and gene therapies: Favourable tax reforms and competition for new assets could lead to higher valuations and product deal amounts.
and efficiently tackle reimbursement issues.” Digital connectivity and advanced technologies will drive the delivery of precision health care, creating a connected ecosystem that will raise the following growth opportunities: • AI and cloud deployment in pharma research and development (R&D): Drug discovery and clinical development IT solutions spending to exceed $10 billion in 2018; • Convergence of molecular diagnostics and big data analytics:
Disruptive start-ups with oncology diagnostics portfolio will see high rewards from their investments; • Smart genomics: Research labs jump-start spending on cloud, NGS and big data analytics solutions; • Biopharma partnerships with contract development and manufacturing organizations (CDMO) and contract research organizations (CRO): Outsourcing of bio-analytical and current good manufacturing practice (cGMP) services is expected to reach $8.5 billion in 2018;
Tetra Bio-Pharma collaborates with Germany’s Storz & Bickel to target fibromyalgia market
Tetra Bio-Pharma and Storz & Bickel agree on a co-development to use a specialised medical device known as the Mighty Medic in combination with Tetra Bio-Pharma’s PPP001 prescription drug as well as future indications and a future proprietary cannabinoid formulation. The Mighty Medic is a portable herbal vaporizer for cannabis use approved by Health Canada as a Class 11 medical device and as such, is eligible for reimbursement. The agreement allows Tetra to effectively bring PPP001
to patients suffering from fibromyalgia and other chronic pain conditions, significantly expanding the market potential for PPP001. Fibromyalgia affects about just over half a million adult Canadian residents or 1.7 per cent of the population.1 Fibromyalgia, also known as fibromyositis and fibrositis, is a common chronic disease, characterized by widespread, constant pain throughout the body, sleeping disturbances, and abnormal exhaustion. Most people with fibromyalgia are women, in whom the initial symptoms appear between the ages of 20 and 50. However, fibromyalgia can also affect men, teenagers, and children. Treatments aimed at pain relief are the primary treatment intervention including an increasing use of opioids. Among the most commonly
prescribed medications used to treat fibromyalgia are Lyrica, Cymbalta, Xyrem, and Vimpat. PPP001 is a cannabinoid-based product that is currently being studied in a Phase 3 Health Canada approved clinical trial for advanced cancer pain. Tetra will use a bridging strategy to leverage the clinical data that has been generated with the smokable formulation of PPP001, to expedite the marketing requirements in these indications and reduce the time needed to bring these products to market. The combination product of PPP001 with the Mighty Medic would signify an important economic benefit for patients. According to Dr. Gordon D. Ko, MD, FCFP(EM), FRCPC, PhD, “fibromyalgia is difficult to treat and there is no one “magic bullet”. As practitioners, we have the daunting task of moving our patients from unrelenting neuropathic pain to improved function and a better quality of life. The clinical use of medical cannabis in the management of fibromyalgia suggests these patients obtain significant relief. Moreover, the potential to
“While the cell and gene therapy segments have shown potential, pharmaceutical and biotechnology companies continue to invest in the promising segments of oncology, cardiovascular, diabetes, and immunology. These segments not only have high unmet needs but also have high acceptability across regions,” says Lal. “Overall personalization, decentralization, collaboration, and prevention are amplifying precision health, thereby shaping the future business models and the workforce of life sciences companies.” To see this story online visit https://laboratoryfocus.ca/megacorporations-to-propel-the-lifesciences-industry-towards-1-5-trillionmilestone/
reduce opioid use for these patients is a foremost consideration of using the Mighty Medic to deliver the cannabinoids and terpenes contained in PPP001 by inhalation,” adds Dr. Ko, who is the medical director, Fibromyalgia clinics and Adjunct Lecturer at The Canadian Centre for Integrative Medicine, (Markham, Ontario) and Sunnybrook Health Sciences Centre, University of Toronto. As a prescription drug, PPP001 will be eligible for insurance coverage. In Canada, only Class II, III, and IV medical devices are eligible for reimbursement under provincial health care insurance programs. The companies plan to collaborate on the development of another inhalation cannabinoid-based prescription drug as well as PPP001. The drugdevice combination product will target cancer patients and Tetra will use a similar bridging strategy to minimize marketing requirements and reduce time-to-market. Reference 1. www150.statcan.gc.ca To see this story online visit https://laboratoryfocus.ca/tetrabio-pharma-collaborates-withgermanys-storz-bickel-to-targetfibromyalgia-market/
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news
Health Canada approves Sunovion’s Aptiom to treat partial-onset seizures for patients with epilepsy even more adults living with epilepsy and partial-onset seizures, which can be challenging to manage.” Phase 3 was a double-blind, activecontrolled, non-inferiority study, consisting of 813 adults that were newly or recently diagnosed with epilepsy. They received either Aptiom (800, 1200 or 1600 mg, once-daily) or the active comparator carbamazepine controlled release (CBZ-CR; 200, 400 or 600 mg, twice-daily). All patients were randomized to the lowest dose level of each drug and only escalated to the next dose level if a seizure occurred. Aptiom met its primary efficacy endpoint of non-inferiority to CBZ-CR. In the primary efficacy analysis, 71.1 per cent of the patients were classified as seizure-free in the Sunovion Pharmaceuticals Inc. announces that Health Canada has approved the use of Aptiom (eslicarbazepine acetate) as monotherapy for partial-onset seizures in adults with epilepsy. All patients who participated in the monotherapy trial were newly or recently diagnosed with epilepsy. Aptiom is now indicated in Canada as monotherapy and as adjunctive therapy for the treatment of partialonset seizures (POS) in adults with epilepsy. “Partial-onset seizures are the most common type of seizures experienced by people living with epilepsy and, given their unpredictable nature, can have a significant impact on their life,” says Eduard Bercovici, M.D., Epileptologist, director of the Southern Ontario Epilepsy Clinic. “This new indication for Aptiom in Canada helps to provide an additional treatment option for health care professionals and patients with partial-onset seizures.” The SNDS approval is supported by data from a Phase 3, double-blind, active-controlled, non-inferiority study in which Aptiom met its primary efficacy endpoint of non-inferiority to the active comparator, carbamazepine controlled release (CBZ-CR). “APTIOM’s approval in Canada as a once-daily, monotherapy treatment option for adults living with partial onset seizures is a significant milestone in Sunovion’s commitment to the epilepsy community and improving the lives of people affected by neurological conditions,” says David Frawley, executive vice president and chief commercial officer at Sunovion. “We are pleased to offer Aptiom as a new monotherapy treatment option for
Aptiom group and 75.6 per cent in the CBZ-CR group within the 26-week evaluation period. The most common treatment-emergent adverse events (TEAEs) reported for APTIOM included dizziness (13.7 percent) and headache (22.9 percent), but Aptiom was generally well-tolerated across the general population of patients. The study (BIA-2093-311) was conducted by Sunovion’s partner, BIAL, a privately held Portuguese research-based pharmaceutical company. To see this story online visit https://laboratoryfocus.ca/healthcanada-approves-sunovions-aptiomto-treat-partial-onset-seizures-forpatients-with-epilepsy/
MedReleaf to collaborate with CFL alumni to study effects of medical cannabis on chronic pain MedReleaf Corp. announces it will collaborate with the Canadian Football League Alumni Association (CFLAA) in conducting an observational study on the benefits of medical cannabis in treating chronic pain and related ailments in retired professional athletes. “We are excited to collaborate with the CFL Alumni Association on this significant observational study with legends in the Canadian athletic arena,” says Neil Closner, MedReleaf CEO. “Through ongoing research, our understanding of the benefits of medical cannabis has grown significantly and we welcome the opportunity to apply our expertise in validating new patient therapies and contributing to positive patient outcomes for this exceptional group of former athletes.” MedReleaf and the CFLAA recognize the opportunity to collaborate on this important study which will bring greater awareness to the potential health benefits of using medical cannabis in the treatment of ailments that include chronic pain. The joint observational study is scheduled to begin in the summer of 2018 with volunteer CFLAA member participants. “The CFLAA has always sought
to make a difference by providing support to our alumni, many of whom experienced medical challenges following their athletic careers,” says Leo Ezerins, CFLAA executive director. “In collaborating with MedReleaf we hope to gain further insight into the benefits of medical cannabis in managing chronic pain – insight that could be invaluable to our almost 2,000 alumni members.” MedReleaf is an R&D-driven company devoted providing top-quality cannabis. They have two state-of-the-art
ICH-GMP and ISO 9001 certified facilities in Ontario Canada, and a third facility currently in development. MedReleaf serves the therapeutic needs of patients seeking safe, consistent and effective medical cannabis for recreational and medicinal use. To see this story online visit https://laboratoryfocus.ca/medreleaf-to-collaborate-with-cflalumni-to-study-effects-of-medical-cannabis-on-chronic-pain/
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Appointments
enGene Inc. appoints Jason Hanson as its new president and chief executive officer as Anthony Cheung Ph.D. steps down but remains at the company as the chief technology officer. Hanson most recently served as president and chief executive officer of Ohana Biosciences, a biotechnology company based in Cambridge, MA. He previously served as executive vice president and chief strategy officer for NuVasive, Inc. and as corporate vice president of General Electric Company and member of the senior executive team of GE Healthcare. While at GE Healthcare he had global business responsibilities for a range of portfolio man-
Jason Hanson
agement, corporate development, legal, compliance, and government relations activities. Prior to joining GE Healthcare, Hanson served as company group chairman and executive vice president at Valeant Pharmaceuticals with responsibility for the company’s Consumer, Ophthalmology, Latin American and Dental businesses, as well as the manufacturing and supply chain, R&D, regulatory and medical affairs teams. Hanson was the executive vice president and chief operating officer at Medicis Pharmaceutical Corporation, where he led R&D and other critical functions and helped build the renowned pipeline of prescription dermatology and aesthetic medicine products prior to its acquisition by Valeant for $2.6 billion. He received a bachelor’s degree from Cornell University and a law degree from Duke University School of Law. St. Joseph’s Health System (SJHS) appoints Dr. Tom Stewart as the new incoming chief executive officer (CEO) of SJHS and Niagara Health. Currently, Dr. Stewart is serving in an integrated system role as joint chief of staff for Ni-
Dr. Tom Stewart
agara Health, where he is also executive vice-president medical, at St. Joseph’s Healthcare Hamilton and at Haldimand War Memorial Hospital. He is set to begin his new role on Aug. 1, 2018. Dr. Stewart is renowned provincially and nationally for his outstanding contributions to the health care system. As well as an award-winning physician, he is also a professor at McMaster University in the department of medicine and formerly at the University of Toronto, where he led numerous innovative clinical trials that were published in leading journals around the globe. He graduated with a Doctor of Medicine with Honours from the University of Ottawa followed by training in internal medicine and adult critical care medicine at the University of Toronto. Following that, he undertook further research training at the University of Paris VI and the University of Toronto. “I am proud to be chosen as the next leader of St. Joseph’s Health System and Niagara Health,” says Dr. Stewart. “We have an exciting opportunity to build on the incredible success that led to the creation of one of the largest health care systems in the country. I look forward to growing the highly regarded and innovative, integrated model of care and our academic mandate, in partnership with leading post-secondary institutions such as McMaster University, Brock University, Niagara and Mohawk Colleges.” Analogic Corporation, a leading provider of advanced imaging technologies to the medical imaging and security industries, welcomes the addition of Mr. Peter Granick as vice president, Operations. Granick will assume overall leadership of Analogic’s global opera-
tions. Fred Parks, president and CEO, comments, “We are delighted to add Peter to our team. His experience leading a world-class global operations team will be a tremendous boost as we continue to look for ways to improve our supply chain and better serve our customers.” Prior to Analogic, Granick served as vice president, Global Supply Chain, Life Sciences Solutions Group at Thermo Fisher Scientific, and brings more than 35 years’ experience in global procurement, operations, supply chain
Peter Granick strategies, and process reengineering. Granick adds, “I am excited to join Analogic and continue to build on the organization’s dedication to operational excellence and commitment to first-class customer service through its unique global footprint.” Clementia Pharmaceuticals appoints an experienced finance executive, Steve Forte, as chief financial officer, effective August 3, 2018. Mr. Forte will replace current CFO, Michael Singer, who will transition from his role effective August 9, 2018. Forte is a senior finance leader with broad experience managing complex, largescale finance environments. His experience includes nearly a decade at Aptalis Pharma Inc., where he was responsible for the overall corporate controllership function of a multinational pharmaceutical company with approximately $700 million in annual revenue, six operating companies, five manufacturing sites and 40 subsidiaries. Forte was responsible for SEC reporting and led the preparation of an SEC S-1 registration statement for a U.S. IPO on NASDAQ prior to the successful sale of the company to Forest Labs for $2.9 billion. He also led the financial due diligence
efforts leading to several acquisitions, including that of a key competitor worth $650 million that transformed the company. Most recently, Forte was CFO of Thinking Capital Financial Corporation, a leading Canadian financial technology firm, where he helped lead the firm through rapid growth and ultimately to a successful sale of the company to Purpose Investments. Forte received his Bachelor of Commerce in Accountancy from Concordia University and is a CPA/ CMA. Clementia Pharmaceuticals Inc. appoints, Shawn Cline Tomasello, to the company’s board of directors. Tomasello has extensive strategic experience in building world class organizations encompassing all aspects of commercial and medical affairs functions. She most recently served as chief commercial officer of Kite Pharma, and was imperative to its acquisition for $11.9 billion by Gilead Pharmaceuticals in 2017. Prior to joining Kite Pharma, Tomasello served as the chief commercial officer of commercial and medical affairs at Pharmacyclics, and was previously the president of the Americas, hematology and oncology for Celgene Corporation where she led the company through
Shawn Cline Tomasello
five successful product launches encompassing eleven indications. Previously, Tomasello served in leading commercial roles with Genentech and other major pharmaceutical companies. She currently serves on the board of directors for Diplomat Pharmacy, Centrexion Therapeutics and Oxford BioTherapeutics. Tomasello received her Bachelor of Science in Marketing from the University of Cincinnati and her M.B.A. from Murray State University in Kentucky.
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feature
B y N i d i a L a u z o n a nd P r . P i e rre C ha u ra nd
State of the art molecular analysis of fingermarks by imaging MS
The use of fingerprints dates back several millennia. Over time, fingermarks, the trace left by fingerprints, have been found in many parts of the world, such as on Paleolithic cave walls, Babylonian clay tablets, Egyptian tomb walls and Greek pottery. Some of these were inadvertently affixed as a natural result of trade, while others were voluntarily affixed, either as a decoration or as a signature to protect the products from counterfeiting. It was not until the 19th century that fingermarks were introduced to forensic science as a means of criminal identification. This advent swiftly replaced the Bertillon system of judicial anthropometry, which is based on the recording of eleven anatomical measures of the head and body, as well as special signs of the criminal (weight, eye color, scars), all accompanied by photographs of the face.
F
or over one hundred years, fingerprints reigned as one of the most trusted identification methods used in forensic investigations. Fingermark recovery is carried out by a crime scene technician using several optical, physical and chemical methods. Initially, a white judicial or laser light source is used to visually inspect the crime scene for fingermarks. Subsequently, one or more physical or chemical revelation techniques are used, followed by photography using various light sources as necessary. In all cases, the ideal forensic enhancement technique must be chosen by the expert in digital trace recovery to avoid destroying the evidence. For this purpose, a better understanding of the chemical composition of a digital trace, namely the fingermark, also referred to as latent fingerprint, is needed. The fingermark results from a transfer of naturally produced substances present on the fingertip or from other body parts such as the face touched by an individual. These substances present on the fingertip originating from skin secretion, including sweat and sebum (endogenous content), are mixed with contaminants from the external environments and microbial communities (exogenous content). Consequently, examining the chemistry of a fingertip can provide a glimpse into the lifestyle and personal habits of the owner, including their general hygiene, diet, med-
ication, personal care products used and locations visited. When applied to forensic investigations, the profiling of a suspect’s lifestyle can lead to a better understanding of the facts of a case, especially when fingermarks found at the crime scene are blurred, dirty or incomplete and do not allow for formal identification. In this regard, the modernization
Figure 1
of chemical analysis technologies led scientists to explore new possibilities to further analyze fingermarks sampled from a crime scene. Mass spectrometry (MS) is an analytical chemistry technique that significantly contributes in many areas of research by providing qualitative and quantitative information on molecular composition. In forensic science, mass spec-
trometry is used to analyze traces for evidence in a police investigation. The scientific interpretation of results contributes to an essential input of information to support police investigations and judicial processes. Since its introduction in forensic science, mass spectrometry has identified various substances found at the crime scene, including explosives, firearm residues, illicit drugs, poisons, pharmaceutical compounds and traces of blood. This area of science is in constant development to counter new stratagems resulting from the ingenuity of some outlaws. Imaging mass spectrometry (IMS) is a molecular imaging technique used to visualize the spatial distribution of molecules across a surface, such as biomarkers in tissue sections for medical research. More recently, this technique has shown to be a powerful
Imaging Mass Spectrometry of Fingermark’s ridges and pores
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feature tool for the analysis of fingermarks, since it combines the detection of numerous endogenous and exogenous substances present at the fingertip’s suspect while creating molecular images of the fingermark, therefore of the fingerprint, allowing suspect identification. After a decade of research in this field, the laboratory of Pr. Simona Francese, at Sheffield Hallam University (United Kingdon), was the first to implement an IMS method into the current fingerprinting workflow of the police authorities. To use this technology here in the province of Quebec, an alternative method was developed in our laboratory for its application on currently available IMS instruments in the city of Montreal. Silver-assisted laser-desorption ionization (AgLDI) IMS was initially developed in our laboratory to profile and image low molecular weight olefin containing molecules from thinly cut tissue sections and is applied here for the chemical analysis of fingermarks. After the deposition of a thin layer of metallic silver (~14 nm) on the fingermarks, we were able to detect numerous endogenous substances, including fatty acids, wax esters, squalene, cholesterol and triglycerides, as well as many exogenous substances, such as a variety polymers and quaternary ammonium compounds found in common personal care products. The silver deposition is carried out using a metal sputtering system in order to provide a homogenous coating across the fingermark for the generation of high spatial resolution molecular images.
Figure 2
Thanks to the conductive properties of silver, this method allows for the analysis of fingermarks left on several nonconductive porous and nonporous surfaces. Indeed, sample conductivity is an essential parameter for IMS analysis using time-offlight mass analyzers, which are commonly found in the Montreal area. However, another important aspect to consider when developing a method for the analysis of fingermarks is their capacity to complement the forensic enhancement techniques already in use by crime scene technicians. Through a collaboration with the fingerprint specialist of the Sûreté du Québec (SQ), we have been able to demonstrate the possibility of generating high quality molecular images of fingermarks first revealed in the SQ laboratory on various nonconductive surfaces, including paper, plasticized cardboard, plastic, as well as fingermarks collected using forensic lifting tape. In this context, the analysis of fingermarks by AgLDI IMS also makes it possible to counter several contrast issues frequently encountered when using optical revealing techniques on multicolour surfaces,
such as cigarette cardboards. Looking at the distribution of individual molecules enables us to generate multiple chemical images of the fingerprint pattern of the suspect independently of the color of the background. Using the chemical specificity of individuals, IMS can also be used to differentiate overlapping fingermarks from two individuals. It can also be used for the detection of overlapping inks in falsified documents. The primary purpose of MS in forensic science is nevertheless to identify substances that can contribute to criminal investigations. In this regard, we also evaluated the potential of AgLDI IMS to detect and map various exogenous substances present in fingermarks, such as those contained in cosmetics, personal care products, illicit drugs (THC, cocaine and heroin) as well as traces of blood. Blood is a highly interesting substance to detect because it can be very useful for the chronological reconstruction of events at the crime scene. Therefore, bloodspecific enhancement techniques are used at the crime scene when blood is suspected to be present. Through a collaboration with criminal identifica-
tion technicians from the Service de Police de la Ville de Montréal (SPVM), we have demonstrated that bloodspecific molecular images can be generated from bloody fingermarks deposited on various conductive and nonconductive surfaces first revealed by common blood forensic enhancement techniques. Beyond the scientific perspectives that can be explored in the laboratory, the continuation of this research is essentially based on the interest of the police forces in implementing molecular imaging in the processes of evidence analysis. We and others have demonstrated that IMS methodology has reached a level of maturity capable of generating high quality molecular images from fingermarks, and we foresee that it will be successfully used in forensic investigations to provide additional information to criminal investigations.
Nidia Lauzon is a Ph.D. candidate at the Chemistry Department of the Université de Montréal in the laboratory of Pr. Pierre Chaurand.
Imaging Mass Spectrometry of revealed Fingermark on a Cigarette Cardboard Pr. Pierre Chaurand (Ph.D. 1994, Université Paris Sud, Orsay, France) is Professor of Chemistry at the Université de Montréal (2009 - present). His expertise’s are in fundamental and analytical mass spectrometry. He is one of the pioneers of the imaging mass spectrometry (IMS) technology. His research interests are focused on the development of new strategies and methods to improve the specificity and sensitivity of IMS with applications in forensic science and clinical biology.
Using the chemical specificity of individuals, IMS can also be used to differentiate overlapping fingermarks from two individuals. It can also be used for the detection of overlapping inks in falsified documents.
To see this story online visit https://laboratory focus.ca/state-of-the-artmolecular-analysis-of-fingermarks-by-imaging-ms/
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feature
B y D e r e k W i ls o n a nd C ri s t i na L e nt o
Advanced Bioanalytical Technologies Accelerate Drug Development in the New Age of
Protein Therapeutics I
n 1986, the FDA approved a new transplant rejection drug that was derived from an intact monoclonal antibody (mAb).1 Although it would be almost a decade before a second mAb drug was approved, this event was the first small step in what has become a foundational shift in the pharmaceuticals industry. From the mid-1990’s, FDA approvals of mAb drugs accelerated steadily, and development pipelines have become increasingly dominated by a widening array of mAb-based candidates. As of 2016, there were over 60 approved mAb-based drugs
with a total market of approximately $89 billion per year.1 It has been estimated that there are more than 550 antibodies in clinical development, approximately 50 of which are in phase III trials.1 The market forces driving the shift towards mAbs are clear: While 90% of prescriptions in the US may be for small molecule drugs, four of the top five earning drugs are mAbs (or mAb-based protein therapeutics) including repeated top-earning drug Humira (Adalimumab) which grossed $18.43 billion in 2017, even while technically offpatent.2 The therapeutic driving forc-
es are equally clear: mAbs use the uniquely high specificity of proteinprotein interactions to tightly and unerringly bind their targets, even in the exceedingly complex molecular environment of the cell. This can have the effect of greatly improving therapeutic power while drastically reducing offtarget effects,
which no doubt contributes to the relatively high success rate for mAbs drugs, more than 15% of which survive from first-in-human clinical trials through to approval.1
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Feature This is not to suggest that mAb drug development is any less challenging than for small molecule drugs. In fact, there are aspects of development, particularly in the pre-clinical phases, where working with biologically manufactured macromolecules can present a broad set of unique challenges. Among these is the very real struggle to determine exactly what the biologically active form of the molecule is, or even if there is a singular form that is responsible for the majority of the activity. While molecular characterization is usually straightforward for small molecules, protein therapeutics have complex, dynamic 3D structures that dictate biological activity, clearance rate and immunogenicity. Furthermore, protein therapeutics are often covalently modified in a manner that is non-uniform and often exceedingly challenging to control during biomanufacturing. The result is invariably a distribution of drug molecules whose on- and off-target effects are determined by the properties of the distribution rather than any one spe-
As of 2016, there were over 60 approved mAb-based drugs with a total market of approximately $89 billion per year.1 It has been estimated that there are more than 550 antibodies in clinical development, approximately 50 of which are in phase III trials. cies. A key factor in overcoming these challenges is the application of powerful new bioanalytical tools that can rapidly provide a detailed picture of candidate protein therapeutics, their molecular, structural and dynamic distributions and their molecular mechanisms of action.3 The ‘Technology-Enhanced Biopharmaceuticals Development and Manufacturing’ program (TBio-DM), led by the Wilson group at York University, is providing leadership in the translation of advanced bioanalytical techniques to drug development. This $1.6M initiative is funded by the NSERC Collaborative Research and
Development program, together with industrial partners SCIEX, Fluidigm Canada and Sanofi Pasteur. Now early in it’s third year, TBio-DM has already delivered on it’s principal aim of accelerating drug development through the application of new technologies in ‘real-world’ projects, often with a direct and substantial impact on business trajectories. Below are three narratives from TBio-DM associated projects and their impact on drug development.
Rapid Characterization of an Avastin Biosimilar The shift to protein therapeutics has
Figure 1
A) A schematic depiction of Native ESI-MS, adapted with permission from ref.4 B) A schematic depiction of Ion Mobility Spectrometry C) A schematic depiction of TRESI-HDX. D) Representations of the type of data acquired by each of the methods.
created additional challenges for generic drug makers, who must now demonstrate biosimilarity for molecules whose structure and activity can be highly sensitive even to minute changes in manufacturing processes, and for which it can be a struggle to demonstrate molecular mechanisms of action. In pre-clinical development of biosimilars, it is necessary to establish biosimilarity not only in terms of conventional biochemical metrics (i.e., Kd; IC50; PK/PE), but also in terms of structural properties, conformational stability and molecular mechanism of action. Apobiologix, a division of generic drug-maker Apotex, has partnered with TBio-DM to develop analytical tools for rapid, facile characterization of an Avastin biosimilar in development. The approach combines newly established advanced bioanalytical methods, like native mass spectrometry (Figure 1A), ion mobility spectrometry (Figure 1B) and technologies unique to TBio-DM, like microfluidics-enabled Time-Resolved ElectroSpray Ionization mass spectrometry with Hydrogen Deuterium exchange (TRESI-HDX, Figure 1C), to provide a detailed picture of the biosimilar and its interaction with the Avastin target, VEGF (Figure 1D, bottom). In the current example, this approach revealed that: (i) The biosimilar exhibited the same epitope on VEGF, indicating the same molecular mechanism of action as the reference product. (ii) The biosimilar glycoforms were somewhat more varied and mature compared to the reference product, corresponding mostly to the presence or absence of one or more terminal galactose residues. Such glycosylation differences can underlie differences in activity and immunogenicity. (iii) The global structure (molecular size) of the biosimilar was essentially identical to that of the reference product, with no significantly populated alternate conformations in the ground state, which can indicate differences in conformational stability, potentially impacting potency and shelf-life. (iv) The manufacturing process delivered good lot-to-lot similarity, with only minor differences in the glycoform profile noted. While these results are not GMP
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Feature release methods (as yet), they can nonetheless contribute to regulatory filings for biologics approvals as orthogonal supportive structural characterization data. They can provide critical insights into the molecular bases of any functional differences that may be observed between the biosimilar and the reference product. The straightforwardness and rapidity of these analyses also allow them to be used in direct support of manufacturing and Quality Control (QC), providing a detailed picture of the biosimilar product at each stage of development.
Small Molecule Drug Development Neurodegenerative Disease While much of the motivation to employ advanced bioanalytical tools comes from the transition to protein therapeutics, the same tools can also substantially enhance the design of small molecule drugs with challenging protein targets. Amyloidotic neurodegenerative diseases (a group that includes Alzheimer’s, Parkinson’s and a host of other neuropathologies) provide a clear example. These diseases are characterized by the intraand/or extracellular accumulation of protein aggregates, in the case of Alzheimer’s corresponding to A-beta and Tau protein. The challenge of amyloidogenic targets is that they are invariably ‘intrinsically disordered’, meaning that they do not adopt a well-defined ‘native’ structure, but rather a weighted distribution of structures (called the ‘conformational ensemble’) that defines their biological function or pathology. Intrin-
sically disordered proteins (IDPs) are notoriously difficult to characterize as they are ill-suited to analysis by the classical tools of structural biology, X-ray crystallography and structural NMR. It is therefore often not possible to apply the conventional strategies of ‘structure-guided’ drug design, nor to (experimentally) uncover molecular mechanisms of action for drug candidates that directly target amyloidogenic proteins. In 2016, Treventis corp., a Toronto company specializing in the development of novel anti-amyloid drugs, partnered with TBio-DM to apply a unique approach for acquiring structural characterization and mechanism of action data candidates targeting the Alzheimer’s implicated Tau protein. Using TRESI-HDX, the TBio-DM team was able to show precisely how Treventis candidates were impacting the conformational ensemble of Tau protein in order to exert their therapeutic effect. These data were specific enough that submolecular features of the drug-altered Tau conformational ensembles could be correlated to drug efficacy in vitro (Figure 2), including the ‘intensity’ of binding to a known amyloidogenic region, which correlated to the ‘steepness’ of IC50 curves, and the extent of global conformational collapse, which correlated to the IC50 value. These molecular mechanism of action data provided a basis for candidate selection, direction for iterative improvements in subsequent rounds of development and, importantly, played a key role in setting the stage for a multimillion dollar research agreement with French pharmaceutical giant Servier.
In pre-clinical development of biosimilars, it is necessary to establish biosimilarity not only in terms of conventional biochemical metrics (i.e., Kd; IC50; PK/PE), but also in terms of structural properties, conformational stability and molecular mechanism of action. Vaccine Potency Assay Reagent Development Unlike their drug-maker counterparts in the pharmaceuticals industry, vaccine developers have always been faced with the challenges of biological production. The active ingredient in vaccines typically corresponds
Figure 2
The impact of three representative Treventis drug candidates on the conformational ensemble of Tau protein, and the connection to IC50. Blue/green indicates regions with reduced exchange in the presence of the drug candidate, suggesting that the protein has adopted a ‘closed’ configuration in these regions. Red/yellow indicates regions with increased exchange in the presence of the drug candidate suggesting that the protein has become adopted a more ‘open’ configuration in these regions. These results allow Treventis to determine which sub-molecular properties of their candidates are required to induce which effects on the conformational ensemble, accelerating the design of subsequent generations of candidates.
to one or more protein (or immunogenic carbohydrate) from the target pathogen, which is produced by the pathogen itself in host cells or by other organisms using a recombinant expression system. As a consequence, vaccine developers can in many cases be viewed as ‘early adopters’ for many of the bioanalytical techniques that will be needed to support mAbbased drug development. One clear example of this is the translation of mass spectrometry-based proteomics techniques (developed largely in academia in the 1990’s) to detect and quantify low-level protein contaminants from pathogen or host cells in formulation. Structural assays to reveal the conformational properties of the antigen and, if possible, the complex with mAbs raised against it, have largely relied on classical techniques like X-ray crystallography and structural NMR. Another important aspect of preclinical vaccine development is the ability to predict potential antigen ‘potency’ in terms of the extent to which it presents surfaces that can be tightly bound by mAbs generated in the patient. This is typically achieved by western blot, which can be carried out in a high throughput manner, but is relatively uninformative in the sense that it provides no information about the nature of the complex
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feature References. 1. Carter, P. J. & Lazar, G. A. Next generation antibody drugs: pursuit of the ‘high-hanging fruit’. Nat. Rev. Drug Discov. 17, 197– 223 (2018). 2. Souriau, C. & Hudson, P. J. Recombinant antibodies for cancer diagnosis and therapy. Expert Opin. Biol. Ther. 3, 305–318 (2003). 3. Deng, B., Lento, C. & Wilson, D. J. Hydrogen deuterium exchange mass spectrometry in biopharmaceutical discovery and development - A review. Anal. Chim. Acta 940, 8–20 (2016). 4. Liuni, P. & Wilson, D. J. Understanding and optimizing electrospray ionization techniques for proteomic analysis. Expert Rev. Proteomics 8, 197–209 (2011).
Figure 3
A) Conventional HDX workflow. B) Diphtheria toxin (left) and epitope mapping HDX analysis for two protective antibodies against diphtheria toxin (right), revealing the location(s) of antibody binding and distinct mechanisms of inactivation.
The challenge of amyloidogenic targets is that they are invariably ‘intrinsically disordered’, meaning that they do not adopt a well-defined ‘native’ structure, but rather a weighted distribution of structures (called the ‘conformational ensemble’) that defines their biological function or pathology. (e.g., precisely where and how binding is occurring) and is only weakly quantitative. Western blots can be complemented by surface plasmon resonance (SPR) or ForteBio® assays to acquire dissociation constants for mAb/antigen complexation, and/ or to identify the location of binding through scanning of antigen peptides. However, this approach is labour intensive and often fails, especially when the mAb binds to an antigen surface that corresponds to discontinuous sequence of amino acids (i.e., when there is a ‘conformational epitope’). Sanofi Pasteur has partnered with TBio-DM to implement hydrogendeuterium exchange (HDX)-based epitope mapping in pre-clinical vaccine development. This approach uses a heavily automated system that is commercially available from Waters Corp. to define the interaction surface for antibody/antigen interactions.
The advantage of this approach is that it is relatively facile, and yet can provide a highly detailed picture of linear and conformational epitopes with moderate throughput. Consequently, it is possible to analyze panels of antibodies for those that might be expected to have ‘protective’ (i.e., inactivating) interactions with the antigen, and to simultaneously determine the mechanism of protection. As an example, Figure 3 illustrates a TBio-DM HDX analysis for two protective antibodies against diphtheria toxin (DTx), revealing distinct mechanisms of inactivation. This information is crucial to reagent (mAb) selection for vaccine potency assays, and can ultimately inform antigen selection for incorporation into vaccine products.
Outlook The ongoing transformation of drug development pipelines to favor pro-
tein therapeutics over small molecules is driving an increasingly pressing need to translate advanced bioanalytical technologies to all stages of drug development in industry. To the companies that invest in the necessary technologies and expertise will go the competitive advantages of substantially accelerated pre-clinical development, knowledge-enhanced decision-making for advancement of early-mid-stage candidates, better control over manufacturing and production QC and, as more sophisticated assays are validated by regulatory agencies, ultimately more success in regulatory filings. Programs like TBio-DM at York university provide a critical bridge for drug developers at all stages, from emerging biotechs to multi-nationals, allowing low-risk, low-cost access to advanced – and sometimes entirely unique – bioanalytical tools that will have a direct impact on business trajectories.
Derek Wilson is a Tier II Research Chair in Molecular Mechanisms of Disease at York University, Director of the Center for Research in Mass Spectrometry and the “Technology Enhanced Biopharmaceuticals Development and Manufacturing” (TBio-DM) initiative.
Cristina Lento is an expert in protein chemistry, bioanalytical mass spectrometry and is laboratory manager of the Wilson Group.
To see this story online visit https:// laboratoryfocus.ca/ advanced-bioanalyticaltechnologies-accelerate-drugdevelopment-in-the-new-ageof-protein-therapeutics/
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Feature
B y Je r r y F i r e ma n
Technology for
Corn Ethanol Measurement Validated Using DOE
T
he primary activities of Stites & Associates, LLC, (SALLC), a technology development and improvement company working in a wide variety of energy applications, include: setting up labs and experiments, evaluating data, evaluating lab operations, and evaluating and improving technologies. Ron Stites, managing member of SALLC, was formerly the director of research for Range Fuels and holds several three patents in alternative fuels. SALLC has a research laboratory in Brighton, Colorado that performs gas chromatography/mass spectroscopy (GC/MS), Cyclic Voltammetry and Optical Microscopy. Stites believes that combining excellent analytical work with diligent research and “outside-the-box” thinking makes it possible for the company to evaluate existing technology as well as gain insight into the best ways to try to improve a technology. “Often this combination results in learning how to make non-obvious improvements that can result in real breakthroughs,” says Stites. In this application, a supplier of industrial equipment wanted to market an existing product to companies producing ethanol from corn. There was anecdotal evidence indicating that the device increased ethanol yield. However, prior to marketing the device the company wanted to find the best operating conditions and determine what performance ethanol producers could expect from the product. The industrial equipment supplier contracted with SALLC to develop a method for measuring how well the supplier’s product improved performance to the corn ethanol market. The first steps were determining the right measurement method and finding a way to preserve samples in order to prevent their deterioration during shipment from plant to lab. After an accurate measurement method had been identified, the next step was running a number of tests to gain insight into what factors affected sample preservation. Finally, researchers performed a design of experiments (DOE) that quantified the effects of these factors, singly and in combination, on sample preservation. The sample preservation method identified by the DOE has to date functioned well on hundreds of samples from ethanol plants across the Midwest.
Producing Ethanol from Corn The process of producing ethanol from corn begins with soaking the grain in water. This makes it easier to separate the components on the corn kernel. The corn is then milled
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feature Figure 1
DP measurements of maltodextrin (HPLC results)
to produce a corn flour that is slurried with water and a heat-stable enzyme - α-amylase - is added. This slurry is cooked in a process known as liquefaction. which uses heat and mechanical shear to break apart starch molecules into small fractions. After liquefaction the corn slurry, also known as corn mash, is cooled to approximately 30oC and glucoamylase enzyme is added to complete the breakdown of starch into glucose. In the fermentation step, yeast is added to the corn mash to convert the simple sugars to ethanol. The breakdown of starch to glucose takes place in two steps, from starch to maltrodextrins and then from maltodextrins to glucose. The equipment supplier hypothesized that its product could hasten the breakdown of starch to glucose while also using less enzymes. The goal was also to minimize “burning,” a phenomenon that occurs when shorter-chain sugars react together and with proteins to form polymers that yeast cannot digest. In fact, some of the polymers are even toxic to yeast. This process is monitored by analysis of the degree of polymerization (DP) of the maltodextrins. But measuring maltrodextrins is challenging. The key difficulty is that the analysis requires specialized techniques that are not available in the plant so samples must be sent to the lab. But the samples are not stable so the breakdown of starch continues during transport.
Increasing Sample Preservation Time There are roughly 200 ethanol plants in the U.S. Midwest and samples would need to be shipped to the SALLC lab from any plant that tried the new product. Stites recognized that he needed to find a way to stop the reaction for 72 hours, approximately the time it would take for a
Table 1
sample to be shipped to the lab and tested. Samples from the actual production process were too inconsistent to be used for creating a sample preservation process. Stites found that spray-dried maltodextrin provides a consistent starting material to evaluate methods of stopping the reaction. Researching the enzymes, he found that they did not work well at low pH and low temperatures. So
he lowered the pH by adding sulfuric acid, cooled the samples and held them at shipping times from 24 to 72 hours before testing. A pH of 2 appeared to prevent the enzymes from working. Stites reviewed published literature and found a new method based on high performance liquid chromatography (HPLC) that could accurately measure maltodextrins. He set up equipment in the SALLC lab to perform the method. The HPLC method easily detected changes in sugar concentration (see Figure 1). Once this preliminary work was completed, the actual DOE was relatively simple. Earlier, while working at Range Fuels, Stites had begun using Design-Expert® software from Stat-Ease, Inc., Minneapolis, Minnesota, after a colleague at the company spoke highly of the program. Stites received training from Stat-Ease and began using the software at Range. ”I am not a statistician, I am an experimenter,” Stites says. “Design-Expert fits my needs because it is designed for use by subject matter experts who are not necessarily experts in statistical methods. The software walks users through the process of designing and running the experiment and evaluating the results. Stat-Ease also provides very good support. I contact them not only for questions about using the software, but also to check out my statistical thinking and they have always been very helpful. I love working with them.”
Factors and results from DOE used to explore sample preservation methods
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Feature Figure 2
As shown in this half-normal plot, DOE proved the pH results to be significant
DOE Methods and Results The factors, runs and results for the preservation DOE are shown above in Table 1. The %<DP9 in the sample at the beginning of the experiment is 25.00%. The response %<DP9 shows how much this value increased during the experiment. The results show that the most important factor is pH. All of the other factors had minimal effects, although incubation at 60oC
made things slightly worse by speeding up the reaction. The best results were observed when pH was adjusted to 2.0 and storage was maintained at 3oC. With this method, the DP increased to only 25.04% after 48 hours. This is less than the standard deviation of the HPLC method and this method can be easily performed in the field. Next Stites needed to determine
The results show that the most important factor is pH. All of the other factors had minimal effects, although incubation at 60oC made things slightly worse by speeding up the reaction.
whether or not these results are significant or if they could have been achieved by chance. Factorial design analysis uses the half-normal plot to identify significant effects. The orange and blue rectangles on the halfnormal plot in Figure 2 show the effects, positive and negative respectively, and the position of these rectangles reflect the relative size. The further the factor effects are from the
line near zero, the more likely they are to be significant. In this case, the factor effects of variable A, which is pH, are significantly greater than the variation between the insignificant effects, demonstrating the statistical significance of the experiment. The method used for run 6 was adopted and worked very well from the beginning for samples from the slurry and liquefaction steps. Some strange results occurred with the fermentation samples – most of the carbohydrates had disappeared. Stites discovered that the sulfuric acid was interfering with the HPLC measurements. The method was modified to remove the sulfuric acid before analysis by treatment with barium hydroxide and filtering. Since this modification was made, the method has successfully worked with samples from all three steps. Ethanol plants from throughout the Midwest shipped hundreds of samples, which were then used to evaluate the performance of the new product. Some plants found significantly better results and others did not see significant improvements. “The DOE was clearly successful in its ability to identify and validate a measurement method that enabled us to accurately evaluate the performance of a new product in a large number of plants under a wide range of operating conditions,” says Stites.
Jerry Fireman is a technology writer who specializes in writing about computer aided design (CAD), 3D printing, computer-aided engineering (CAE), the Internet of Things (IoT), electronic engineering, pharmaceutical research and manufacturing, test and measurement and a variety of other topics.
To see this story and all the figures online visit https:// laboratoryfocus.ca/technology-for-corn-ethanol-measurement-validated-using-doe/
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New Products Analyzer Technologies The two Spectro products that are inductively coupled plasma optical emission spectrometry (ICP-OES) and energy dispersive X-ray fluorescence (XRF) technologies have seen dramatic improvements in recent years. Spectro flagship instruments showcase the advances that this field has seen with the Spectro Arcos ICP-OES analyzer and the Spectro XEPOS ED-XRF analyzer. Spectro Arcos high-resolution ICP-OES analyzer represents the pinnacle of productivity and performance as the first and only spectrometer featuring the fast, convenient selection of axial plasma or radial plasma observation in a single instrument — without any optical compromise. The analyzer delivers dramatically improved sensitivity, stability, and precision — at lower operating costs. Spectro Xepos ED-XRF spectrometer represents a quantum leap in ED-XRF technology, providing breakthrough advances in the multi-elemental analysis of major, minor, and trace element concentrations. New developments in detection deliver quality sensitivity and detection limits and gains in precision and accuracy.
http://icp-oes.spectro.com/new-analyzer-software
Advanced Cells-to-CT Kits
New Invitrogen Fast Advanced Cells-to-CT kits offer researchers the ability to skip RNA purification entirely by going straight from cultured cell samples to measuring relative gene expression by real-time RT-PCR (RT-qPCR). And for flexibility, Cells-to-CT kits are available in both TaqMan and SYBR Green formats. These kits provide a complete workflow for RT-qPCR analysis directly from cultured cells without RNA purification. Featuring a unique method for lysing cultured cells while removing genomic DNA (gDNA) and preserving RNA integrity, the kits contain reverse transcription (RT) reagents for cDNA synthesis and either TaqMan or SYBR Green master mixes for real-time PCR analysis. The new TaqMan Fast Advanced Cells-to-CT Kit utilizes an improved reverse transcriptase enzyme that is more sensitive to detect abundant or rare transcripts. This makes it possible to perform expression analysis directly from cultured cells without RNA purification. The new SYBR Green Fast Advanced Cells-to-CT Kit is a fast, easy, and robust method to go directly from cultured cells to real-time PCR results. The kit is powered by Applied Biosystems AmpliTaq Gold DNA Polymerase, LD (Low DNA), which provides the highest levels of specificity with standard real-time PCR. The Cells-to-CT Stop Solution allows for greater sample input in the downstream RT and qPCR reactions, greatly increasing the sensitivity for detecting rare targets. Both TaqMan and SYBR Green Fast Advanced Cells-to-CT Kits use the Stop Solution, providing excellent sensitivity with a very fast workflow. These kits are approximately 48 times faster than most high-throughput RNA purification kits. Fast Advanced Cellsto-CT kits can process 96 or 384 wells at once; the experimental workflow only takes about 10 minutes for a 96- or 384-well plate. Products from other suppliers use low pH and dilution in their workflows, which restricts how much cell lysate can be used in downstream steps.
www.thermofisher.com
Pinch Valves Emerson is expanding its ASCO Series 045 solenoid pinch valves to include a new 3-way format with one of the broadest offerings of fluid-handling and control products for the clinical diagnostic market. Previously, the series included only 2-way normally-closed (NC) and normally-open (NO) options. The ASCO Series 045 2-way and 3-way valves effectively control the flow of aggressive, high-purity fluids in clinical diagnostic technologies and bioinstrumentation, including hematology platforms. Valve design feature hermetic separation of the control mechanism and fluid—preventing particulate contamination and ensuring sample and reagent purity. The series comes in a variety of body sizes, meets all relevant CE directives and is RoHS compliant. Emerson will showcase its ASCO Series 045 solenoid pinch valves and other fluid-handling components, including miniature liquid isolation, proportional and general service valves, fittings, manifolds, tubing and pressure regulators, at the upcoming 70th AACC Annual Scientific Meeting & Clinical Lab Expo.
http://www.emerson.com/en-us/automation/asco
Refrigeration Units Thermo Scientific releases the TSG Series powered by Phononic solid-state refrigeration technology. It has been designed to meet the challenging needs of health care, research and industrial applications where solid performance is required, in support of breastmilk and medication storage, analyzer kits and reagents, and even cell culture media. The TSG Series brings cutting-edge innovation into refrigeration by incorporating solid-state technology in place of traditional refrigeration methods. They have increased storage space and use less energy outputting almost no heat into the work environment. The refrigerators are available in two sizes that have been designed for under the counter and the countertop. Model specific features: • Remote alarm contacts on TSG505, supporting requirements of research for monitoring • Datalogging and download capability on TSG205 via USB port to support clinical record management requirements Warranty for both products is 5 years on the solid state refrigeration system and 2 years parts and labor Key features of both models: • Solid state refrigeration technology • Whisper quiet operation • Energy Star rated performance meaning lower operating costs • Green refrigerants supporting global initiatives to reduce greenhouse gas emissions • Full alarm set, including hi/lo temperature, door ajar and power fail to support requirements for medical storage
www.thermofisher.com
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Laboratory Focus August/September 2018
Next-generation Sequencing Assay Thermo Fisher Scientific launches its Oncomine TCR Beta-SR Assay, a new nextgeneration sequencing (NGS) solution for immuno-oncology research designed to characterize T-cell clonality in the tumour microenvironment – a key approach to identify potential biomarkers associated with immune response. The Oncomine TCR Beta-SR Assay is designed to specifically interrogate the CDR3 region of the beta chain of the T-cell receptor, a region that is responsible for antigen recognition and important in uniquely identifying T-cell clones. The assay’s utilization of dual barcode indexing also enables identification of low-frequency clones which has the potential for tracking minimal residual disease. Interrogation of data obtained with the Oncomine TCR Beta-SR Assay (SR = Short Read) is optimized for formalin-fixed paraffin-embedded (FFPE), peripheral blood leukocytes (PBL), peripheral blood mononuclear cells (PBMC) or cell-free DNA (cfDNA) samples. One of the features that makes the Oncomine TCR Beta-SR Assay unique is its flexibility in design for translational researchers to use either DNA or RNA input without interference from primer bias while returning results within 48 hours on the Ion GeneStudio S5 Series instruments and Ion Torrent analysis solutions. When coupled with the Oncomine TCR Beta-LR Assay (LR = Long Read), researchers can achieve a comprehensive range of TCR beta sequencing applications, including TRBV allele typing, clonality association to immune response and engineered T-cell monitoring.
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Genetic Testing Kit for Thalassemias Alpha and beta-thalassemia are severe forms of anemia. Alpha-thalassemia is most often caused by deletions in one or both HBA genes, while the major cause of beta-thalassemia is single nucleotide variants (SNVs) in the HBB gene. There are many advantages to this test, which includes: • One simple assay for all thalassemia genetic testing needs • Single tube NGS analysis of mutations causing both alpha and betathalassemia • Detects both SNVs and deletions in the HBA1, HBA2 and HBB genes • Simple, fast and robust NGS workflow – NGS has never been easier • Optimized data analysis software Devyser Thalassemia detects all mutations in HBA1, HBA2 and HBB including SNVs, Indels and CNVs. It is optimized to be used for both large scale mutation screening and advanced second level genetic testing. The fast, simple and robust NGS workflow replaces time consuming multi-step protocols and the need for maintaining several different assays in your lab. Devyser’s proprietary PCR chemistry provides complete and uniform coverage of the targeted regions. It also enables a high level of overlapping amplicon multiplexing, which results in a unique NGS library preparation process with no sample splitting. This simplifies the workflow, reduces the risk of contamination and minimizes the potential for sample mix-up.
www.devyser.com
Liquid Biopsy Solution Exosomics Siena has launched its first proprietary liquid biopsy solutions, SeleCTEV-DNA and SortEV-RNA, that are both designed to aid selective isolation of tumour-derived nucleic acids from complex biofluids such as plasma or serum. Unlike most liquid biopsy methods where total nucleic acids are harvested from blood this technology is based on the isolation and enrichment of tumour-derived exosomes. The new
New Products kits use unique affinity methods to select exosomes from which the DNA or RNA can be extracted. Additionally, the SeleCTEV-DNA kit can be used to isolate circulating cell-free DNA in addition to DNA borne by exosomes. The kit is designed to isolate in one step tumour-originated EVs and exosomes from complex biofluids. It includes an optimized extraction kit for the co-isolation of cell free and EV-DNA. The line features different versions of the isolation kit, including a wide range of sample volumes (from 0.5 to 7 ml) and extraction kit for cf- and EV-DNA. No time-consuming ultracenrifugation step needed, turn-around time is < 3 hr.
www.exosomics.it
Genetic Polymorphisms Detection Kit Medical and Biological Laboratories launches the MEBRIGHT NUDT15 kit for detecting genetic polymorphisms in the NUDT15 (Nudix Hydrolase 15) gene, which can predict serious side effects of thiopurine drugs. Thiopurine drugs are often prescribed to patients with inflammatory bowel disease, leukemia, rheumatic disease, or after an organ transplant, but clinical studies have shown that the drugs can induce adverse effects such as severe leukopenia and alopecia universalis within about eight weeks. A strong correlation has also been demonstrated between the development of serious adverse effects of thiopurine drugs and polymorphism in NUDT15. Using real-time PCR, the kit detects polymorphisms at codon 139 of NUDT15 in DNA from a patient’s blood. The kit requires a processing time of two hours and simultaneously measures multiple samples. The kit was co-developed with Tohoku University, and was approved as an in vitro diagnostic test by Japan’s Pharmaceutical and Medical Device Agency in April.
http://www.mbl.co.jp/e/
Evaporator Genevac HT series evaporators are the ideal solution for parallel evaporation bottlenecks in high throughput and production laboratories having high performance and high sample capacities. The all new Series 3 HT evaporator range from Genevac represents the ultimate in solvent removal technology. They incorporate a new high-performance vacuum pump, the latest touchscreen technology and a sleek ergonomic design that will make the evaporation process effortless. New intuitive touchscreen controls enhance monitoring and review of the whole evaporation process. For popular solvent removal protocols pre-set “Press & Go” methods makes operation easy and productive even for occasional users. Simplified manual and automatic programming means even the most complex multi-stage evaporation methods can be set-up and run quickly and easily. The Series 3 HT evaporator range comes with patented temperature control and bumping prevention built in.
www.genevac.com
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Laboratory Focus August/September 2018 www.laboratoryfocus.ca
app review Instrument Connect
https://play.google.com/store/apps/details?id=com.thermofisher.icma_android&hl=en
Canada in the limelight Machine learning is augmenting human ability and drastically changing possibilities. It is restructuring businesses and rewiring brains for transformative thinking. Whether it be to develop vaccines for deadly diseases or combat climate change, Canada is at the forefront of this monumental shift. Canada has been on the vanguard of machine learning long before it became a popular headline. The growth and brilliant minds from around the country have led the way for Canada to place its mark with AI on the world and build a more promising ecosystem for the future. Across the country, there have been several companies lending a hand in this newer and multi-faceted industry that will reshape history. Among those, innovative researchers are developing imaging devices for skin cancer, diagnostic platforms that analyse natural speech to detect dementia and mental decline, advanced signal analysis to diagnose coronary artery disease, just to name a few, and numerous others that stem to topics unrelated to health care, but keep Canada at the forefront of change. This year Canada had its 31st Canadian Conference in Artificial Intelligence that stands to show how long Canada has been involved with this game-changing technology. Events like this bring together hundreds of leaders in research, industry, and government that provide a melting pot of inquisitive and likeminded people. Artificial intelligence and deep learning have the potential to revolutionize health care, and with Canada as a global frontrunner, we can expect to see exponential shifts in the upcoming years to come.
Instrument Connect remotely monitors instruments in real-time. Stay connected to cloud-enabled instruments and benchtop devices anytime, anywhere, using the Instrument Connect mobile app. With the Instrument Connect remote monitoring app powered by Thermo Fisher Cloud, you can stay connected to any Thermo Fisher Cloud-enabled instruments, including the Applied Biosystems QuantStudio 3 and 5 realtime PCR instruments, and the ProFlex and SimpliAmp end-point PCR devices.
Bio-Rad Events
https://play.google.com/store/apps/details?id=me.doubledutch.bioradeventmanager&hl=en This mobile app allows you to view schedules, explore sessions, and find after parties. You can create your own personal schedule, access location and speaker information. Check-in to sessions, meetings, keynotes, and exhibitor booths. View an entire feed of the event activity, including attendee check-ins, photos, and more.
DNAApp: DNA sequence analyzer
https://play.google.com/store/apps/details?id=bii. seqdatreader&hl=en This is the first android app that allows for the opening and analysis of DNA sequencing files - ab1. It includes handy tools such as “Reverse Complement”, “Jump to”, fast and end scrolling, “Chromatogram adjustments”, amino acid translations, “export to fasta”, and “searching for segment” function. This app works with cloud storage access like Dropbox to your sequencing files.
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My Lab. My Centrifuge. The new Centrifuge 5425 Are you looking for a silent companion who supports you with DNA extraction workflows and cell harvesting? The new Centrifuge 5425 is the latest generation of a more than 50 year history of Eppendorf microcentrifuges.
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