Lab Business March/April 2018

David Suzuki Can insects revolutionize the way we eat and produce food?

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Gene therapy may offer optimism for treating rare eye diseases

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CONTENTS 11 Save the Planet; Eat an Insect

By David Suzuki

Adding bugs to our diet could help us minimize our environmental footprint.

Rebuilding Sight

The Price of Plastic

By Jeff Elliott

By Melissa Wallace

Manufacturers are driving up the value of their labware products by altering their surface properties.

A new research trial at the University of Alberta’s MacDonald Lab seeks to halt a rare inherited retinal disorder in its tracks.

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standards editor’s note 5 Canadian news 6 worldwide news 7 Lab ware 20 moments in time 22

Do the flip!

Discover the hope gene therapy is offering those with genetic disorders.

HUMAN RESOURCES

Approaching employee benefits programs with a pioneering mindset 15

David Suzuki Can insects revolutionize the way we eat and produce food?

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NEWSMAKER

Bioenterprise’s Dave Smardon on being an ecosystem builder 19

MOMENTS IN TIME

Howard Borden Newcombe paves way for advances in molecular genetics 22

MARCH/APRIL 2018

Polymer Labware THE DEFINITIVE SOURCE FOR LAB PRODUCTS, NEWS AND DEVELOPMENTS

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Altering surface properties improves test results

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suzuki matters

HOPE of

Gene therapy may offer optimism for treating rare eye diseases

GENE THERAPY New tools continue to unlock doors in the burgeoning field

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Paying for the Golden Goose S

cientists are wary of using the word cure. Does a gene therapy that could potentially regrow damaged nerves in a patient with a spinal cord injury constitute a cure? What do you call a treatment that halts the progression of a degenerative retinal condition by inserting healthy genetic material into the retina? Surely “cure” would be the most accurate description.

Not so fast, scientists caution. “Technically, a cure means you have eliminated all possibility of risk from the disease,” says Aneal Khan, medical geneticist and lead researcher of a University of Calgary human clinical trial testing a gene therapy treatment for Fabry disease (see Bio Business, page 10). There’s no way to tell whether such gene therapies will be a long-term solution to rare genetic disorders like Fabry, Khan says.

The promise of gene therapy for those with genetic disorders is tantalizing; it could turn their diagnoses from death sentences into manageable conditions, maybe even eradicate these conditions altogether. But none of that will happen without private dollars, Khan says. In the case of the Fabry treatment, U.S. company Avrobio has taken the baton from the University of Calgary researches and will now garner investments to pay for its Phase II and Phase III trials, take it through its development stages, and apply for regulatory approval around the world.

“In Canada we think there’s a golden goose which sometimes lays the egg for funding treatments and access to all these health care technologies, but there isn’t,” Khan says. “We live in a capitalist society and any new technology like this that is going to go forward, somebody has to invest dollars.”

The field of gene therapy is still very young.

Trials have taken place with animal models and are just now moving to humans, says Ian MacDonald, Professor Emeritus at the University’s Department of Ophthalmology and Visual Sciences, and lead scientist for Canada’s first human gene therapy trial for choroideremia (see page 11). “We don’t know exactly what the outcomes will be, but we perceive it will be favourable,” MacDonald says. “Maybe one day, gene therapies will be used for both rare and Hermione Wilson common conditions.”

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Canadian NEWS How do Bone Marrow and Blood Cells React in Space?

In the context of the next SpaceX resupply mission scheduled in April, NASA held a teleconference in March to discuss scientific experiments performed on the International Space Station (ISS). Researcher Dr. Guy Trudel discussed the Canadian experiment MARROW, conducted in collaboration with the Canadian Space Agency, which is examining the effect of space on bone marrow and blood cells. Canadian Space Agency astronaut David Saint-Jacques will be participating in MARROW.

Detecting and Destroying Biofilms Wins Innovation Award

New technology that is able to detect and destroy biofilms on surfaces in food processing plants has been selected as the 2016 award winner for Productivity and Technology Advancement by CTAQ, the premier food processing council of Quebec.

Each year, CTAQ recognizes the industry's top efforts to develop and/or launch projects that meet the criteria for innovation in advancing food safety.

This year’s award was given to Sani Marc Food & Beverage Division for BioDetect and BioDestroy.

Government Investing 5.2M in Science and Innovation at the University of Saskatchewan

While visiting the University of Saskatchewan, the Honourable Ralph Goodale, Minister of Public Safety and Emergency Preparedness, highlighted recent federal investments of $5.2 million for scientific research at the university. The money will be divided among six health research projects as well as a study on the welfare of farmed pigs.

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March/April 2018 Lab Business

CNL Issues RFP for Construction of $370-Million Lab Complex

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anada’s national nuclear laboratory invites proposals for the design, engineering and construction of a major nuclear research laboratory, the Advanced Nuclear Materials Research Centre. Canadian Nuclear Laboratories (CNL), Canada’s premier nuclear science and technology organization, announced that it has issued a Request for Proposals (RFP) for the design and construction of its Advanced Nuclear Materials Research Centre (ANMRC), a $370-million modern laboratory research complex that will serve as the backbone of CNL’s research and development infrastructure and a major facility in the delivery of its nuclear science and technology programs. The RFP is now available for review on Merx, an online tendering service.

Scheduled to begin construction in 2019, CNL’s ANMRC will be one of the largest active research facilities ever to be constructed in Canada, and will enable world-class research in nuclear energy, public health, environmental stewardship and global security.

“The revitalization of the Chalk River Laboratories campus is designed to position CNL as a global leader in nuclear science and technology, transforming the site into a world-class and sustainable national nuclear laboratory,” says Mark Lesinski, CNL’s President and CEO. “The ANMRC is at the centre of this transformation, and our vision for the future of the site and the nuclear industry in Canada. Once complete, the facility will serve as a state-of-the-art laboratory complex that will allow us to grow our research programs, penetrate new international markets, and add capabilities to better meet the needs of our federal, academic and commercial customers.”

As the largest single capital investment in the revitalization of the Chalk River campus, the ANMRC will consolidate key capabilities from a number of aging facilities that are scheduled for decommissioning. The ANMRC will feature new shielded facilities that will enable post-irradiation examination of small modular reactor (SMR) and next-generation nuclear fuels, and glovebox facilities to support the development of advanced fuel fabrication concepts. The complex will also include materials storage bays which will simplify the transportation of radioactive material on site, improving work efficiency at the Chalk River campus. Overall, services provided by the ANMRC will be critical to the life extension and long-term reliability of existing reactors, including Canada’s fleet of CANDU nuclear power reactors and other designs deployed around the world.

Construction of the ANMRC is part of a 10-year transformation of the Chalk River Laboratories site, funded through a $1.2-billion investment from Atomic Energy of Canada Limited on behalf of the federal government, to modernize the campus. This includes the revitalization of essential site infrastructure, the decommissioning of aging buildings and a significant investment in new, world-class science facilities.

To learn more about CNL’s long-term strategy and revitalization of the Chalk River Laboratories, visit www.cnl.ca/vendors.

Worldwide NEWS

What Do Spacecraft, Newborns and Endangered Shellfish Have in Common?

Spacecraft assembly facility. Photo credit: NASA – JPL – Caltech

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new technique can reveal previously undetectable bacteria in places where they aren’t wanted — such as a spacecraft assembly facility, a neonatal intensive care unit and an abalone rearing centre.

Researchers at University of California San Diego School of Medicine have developed a microbial detection technique so sensitive that it allows them to detect as few as 50 to 100 bacterial cells present on a surface.

The technique was validated by sampling hundreds of surfaces in three different environments: the Spacecraft Assembly Facility at NASA’s Jet Propulsion Laboratory at California Institute of Technology; the neonatal intensive care unit (NICU) in Jacobs Medical Center at UC San Diego Health; and an endangered white abalone rearing facility at National Oceanic and Atmospheric Administration (NOAA)’s Southwest Fisheries Science Center in La Jolla, CA. Details of the technique, called KatharoSeq, are published March 13 in the journal mSystems. KatharoSeq has already revealed new insights about the three testing sites that could help optimize how the Mars 2020 Rover is assembled, how bacteria are tracked in hospitals, and how endangered white abalone are raised and returned to the wild.

The Spacecraft Assembly Facility at Jet Propulsion Laboratory is where NASA builds spacecraft and machinery to be launched into space. Engineers and staff must account for every biological organism sent into space in order to prevent contamination of other planets.

Senior author Rob Knight, PhD, professor of pediatrics and computer science and engineering, and Director of the Center for Microbiome Innovation at UC San Diego, and his team wondered if KatharoSeq might be able to detect microbes in what is thought to be a sterile facility. Of the three sites tested, the Spacecraft Assembly Facility had the lowest microbial diversity. But bacteria were still present – KatharoSeq detected 32 types.

Knight’s team will now work with Jet Propulsion Laboratory staff to create a map of the microbes living in the facility over the next six months, including the Mars 2020 Rover.

Their goal is to send a sterile rover to Mars.

The NICU at Jacobs Medical Center at UC San Diego Health is relatively new – the 245-bed advanced specialty hospital opened in La Jolla in late 2016, about four months before samples were collected for this study.

In this study, the NICU samples contained more bacterial species than the Spacecraft Assembly Facility, but fewer than the white abalone rearing facility.

In the future, co-author Jae Kim, MD, Associate Clinical Professor of Pediatrics and Nutrition Medical Director of the Supporting Premature Infant Nutrition Program, and his team hope to investigate probiotic interventions in the NICU. In this study, KatharoSeq detected a diverse microbial community in the white abalone tanks. The researchers found many types of marine bacteria. Knight’s team hopes to work with the rearing facility to determine the optimal microbial make-up of the white abalone and its surrounding environment to improve transplantation success.

Read the full study at http://dx.doi.org/10.1128/mSystems.00218-17

Forum Focuses on Regenerative Medicine and Cell Therapy

Promoting interdisciplinary exchange between science and industry on the topics of regenerative medicine and cell therapy, Sartorius held its second Research Xchange Forum in March in Germany. Experts reported on new developments in cell therapy. Speakers included Dr. Priscilla N. Kelly, editor of the journal Science, and Dr. Stefan W. Hell, the winner of the Nobel Prize in Chemistry 2014 and a professor at the Max Planck Institute for Biophysical Chemistry.

Copley Scientific Appoints Local Expert as COO

Following the recent expansion of its UK headquarters, Copley Scientific announced a new addition to its senior management team, in response to a record year for sales and the continued success of the business. Richard Postlethwaite, formerly Senior Operations Manager, brings local and global pharmaceutical industry experience to the already fast-growing company.

Postlethwaite has been appointed COO and will oversee and optimize all aspects of the business.

High-Risk Industries Uplift Demand for Eye and Face Protection Equipment

High-risk industries are increasing demand for eye and face protection equipment in North America, says Frost & Sullivan. The latest analysis finds that the industry generated revenues of $1.1 million in 2017, and is expected to grow at a compound annual growth rate of 2.9% to reach $1.3 million by 2022.The fastest-growing segments are expected to be eye and face protection, followed by head and ear protection, due to the rise of increased worker safety regulations. www.labbusinessmag.com

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Worldwide NEWS New Researchers Shape the Future of Scientific Information and Innovation

CAS, a division of the American Chemical Society, recently announced the participants selected for the 2018 CAS SciFinder Future Leaders program. The international PhD students and postdoctoral researchers were chosen for their academic accomplishments and the scientific merit of their research. They will visit CAS headquarters this August to collaborate on new initiatives to support the scientific community, including CAS’s innovative workflow solution, SciFinder. The program connects fledgling scientists with leading researchers.

Storage System Accelerates Collection of Data-Intensive Research

DataDirect Networks (DDN) recently installed its GRIDScaler GS7KX parallel file system appliance with 1PB of storage at Harvard University’s Faculty of Arts and Sciences Research Computing (FASRC). The installation has increased the speed of collecting images detailing synaptic connectivity in the brain’s cerebral cortex. The new file system will be able to keep pace with thousands of users – university researchers and affiliated organizations – who create vast amounts of data from complex research equipment that can strain storage capacity.

Methods for Evaluating Soil Health to be Standarized

The Soil Health Institute has released initial methods for cooperating scientists to use in assessing soil health indicators, as they move toward standardization of soil health measurements across the continent.

The lack of widely applicable measurements and methods for assessing soil health are significant barriers to adopting soil health practices and systems. A panel of experts from the U.S. Department of Agriculture, several universities and the private sector developed consensus on how each indicator should be measured.

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March/April 2018 Lab Business

MilliporeSigma Collaborates with Washington University on Restoring Healthy Gut Microbiomes

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illiporeSigma, a leader in genome editing, recently announced a collaboration with Washington University that could lead to opti m izi ng nutr itiona l supplements for the restoration of a healthy gut microbial community or microbiome.

The research aims to determine the differences between gut bacterial communities in healthy and malnourished children, and to identify what features of healthy intestinal bacteria are critical for supporting healthy growth. From there, nutritional approaches to restore a normal microbiome can be developed and optimized, as nutritional interventions to date have failed to solve the problem.

The two-year collaboration will employ MilliporeSigma’s CRISPR genome-editing technology in research studies by Dr. Jeffrey Gordon, director of the Edison Family Center for Genome Sciences and Systems Biology at Washington University.

“Development of the gut microbiome is disrupted in severely malnourished children, leaving them with immature communities compared with healthy children,” says Udit Batra, CEO, MilliporeSigma. “Our collaboration will focus on repairing and reconstituting a normal microbiome in malnourished children. We will continue to form collaborations with the global scientific community to explore how to develop exciting new treatments for many diseases.”

CRISPR-based genomic scissors have unveiled new possibilities in medicine and biotechnology. Similar to a word processing program that finds, deletes and replaces words or letters, these CRISPR RNA-protein complexes search for certain DNA sequences in a cell, cut them and allow the cell to paste in new DNA information. MilliporeSigma, together with Dr. Gordon’s group, will use its CRISPR genome scissors in this collaboration to modify the sequence of DNA in microbes cultured from human gut microbiome samples. The results will help researchers obtain essential, new information about the microbes’ functions and nutritional needs.

“Our shared goal is to apply gene-editing technology to further understand the mechanisms by which beneficial human gut microbes promote healthy growth in children,” says Gordon. “By marrying this technology with our preclinical models, we can decipher how gut microbes become established in the developing gut, what nutrients are necessary to sustain those microbes and how gut microbial communities influence muscle and bone growth, maturation of our immune systems and metabolic health.”

Development of a healthy gut community is linked to the healthy growth of infants and children. “Results obtained from this collaboration should aid our ongoing efforts to devise new, safe and culturally acceptable ways to repair the developing gut communities in malnourished children or children at risk of malnutrition. This knowledge will facilitate development of new types of microbiota-directed foods, composed of naturally occurring ingredients, that increase the representation and beneficial functions of naturally occurring bacterial strains in the immature gut communities of these children,” Gordon says.

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Suzuki MATTERS

By David Suzuki with contributions from Ian Hanington

SAVE THE PLANET; EAT AN INSECT P

Dr. David Suzuki is a scientist, broadcaster, author, and co-founder of the David Suzuki Foundation.

Ian Hanington is Senior Editor, David Suzuki Foundation.

Learn more at www.davidsuzuki.org.

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eople sometimes get bugged by insects, but we need them. They play essential roles in pollination, combatting unwanted agricultural pests, recycling organic matter, feeding fish, birds and bats, and much more.

They’re the most numerous and diverse animals on Earth and form the base of many terrestrial and aquatic ecosystems.

Our admiration for these critters goes beyond their ability to adapt, their fantastically diverse colours and shapes, and their accomplishments that create dramatic impacts on our world’s functioning.

Could the same six-legged creatures that form the backbone of ecosystem services also help minimize humanity’s environmental footprint?

Could insects revolutionize the way we eat and produce food?

We will be nine billion people on Earth in 2050.

To feed that many, we should double food production, according to the UN Food and Agriculture Organization. But the way we currently produce food weighs heavily on the environment.

If food were a country, it would rank third behind China and the U.S. as one of the largest greenhouse gas emitters. We eat too much meat, and its production is disastrous from an environmental standpoint. In Canada, we’re struggling to curb water pollution related to corn and soybean production for livestock feed, and aquaculture relies on wild-caught fish to produce meal and oil feedstock.

Although vegetarian diets would suffice to feed humans and drastically reduce our environmental footprint, meat consumption remains a firmly established tradition. But if we bartered beef, pork or chicken for a handful of insects, the environmental impact of our animal-protein intake would drop dramatically. According to the FAO, 18 per cent of all greenhouse gas emissions are linked to animal husbandry. Emissions from

March/April 2018 Lab Business

insect production are negligible in relation to the amount of protein produced. Insects are especially effective at converting their food because they’re cold-blooded and therefore waste less energy to keep warm.

Although few people in Canada have integrated insects into their regular diet, nearly half of us have tasted an insect, according to a survey conducted in Quebec. According to the study, males are significantly more likely than females to bite into an insect like a fly larva!

To reduce our environmental footprint, not everyone needs to incorporate these crisp, delicious arthropods into their diet. Changing the diet of farmed animals could also help. People who may not want to ingest insects themselves would likely have little problem feeding their pets insectcontaining kibble, or eating farmed animals like chickens raised on insect-based feeds. The Canadian Food Inspection Agency has already licensed fly larvae to feed farmed salmon and chickens. (Full disclosure: My suggestion to use insects to feed carnivores like salmon inspired a company in which I have invested that harvests insects grown on food waste.)

Emerging entotechnologies (from the Greek root entomo, for “insects”) bring together applications that focus on what insects do best. For example, organic residues can be fed to fly larvae, which can then be used as livestock feed. Black soldier fly larvae have voracious appetites for fruit and vegetable residues and could help improve the way we handle this high-quality organic waste.

It’s a way to give a second life to stale food, rather than sending it to compost bins or biogas plants.

Considering nearly 45 per cent of fruit and vegetables produced worldwide is wasted, this is not a fringe idea.

After feeding the hungry with the highest quality unsold portions of our food, we could feed our breeding animals with insects raised on organic residues from grocery stores and restaurant kitchens.

With clean organic residues from urban agriculture, we could also feed humans! In Montreal, the Blanc de Gris farm grows grey oyster mushrooms on recovered coffee and brewery residues.

But we can take it even further. Several tonnes of mycelium (akin to the “roots” of fungi mixed with growth substrate) are discarded from the company’s mushroom production each year. A research project by the David Suzuki Foundation, Montreal Biodome, Concordia University, Université du Québec à Montréal and Université Laval is finding out if mealworms could grow on this mycelium.

The first results are promising. In fact, entotechnology and entomophagy (eating insects) research is blooming in Canada.

You can join the research on edible insects by participating in a survey by Concordia University PhD student Didier Marquis. LB

Lab PROFILE

Fighting the Decline into Darkness story by

Melissa Wallace

Through gene therapy, scientists in Alberta are offering a glimpse of hope to sufferers of a rare retinal disease

www.labbusinessmag.com

11

Lab PROFILE

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horoideremia (CHM) once signalled utter despair among patients. Affecting 1 in 50,000 people – 90 per cent of whom are men – this rare, inherited disorder is caused by a genetic defect of the X chromosome which prevents the production of an important protein and eventually kills retinal cells. For people with choroideremia, this meant experiencing a gradual loss of vision starting in childhood and total blindness by the age of 40.

But a research team at the University of Alberta’s MacDonald Lab is hoping they’ve figured out how to stop the process.

In June 2015, the team pioneered Canada’s first human gene therapy trial for eyes with a mission to preserve and potentially restore vision for people with CHM. Six Canadians with CHM who had expressed interest in the clinical trial were selected from 20 individuals. They underwent surgery where a vector (or customized virus) was injected, transferring healthy, genetic material into the retina. Next, patients received regular monitoring and endured a series

(From left to right): The MacDonald Lab team members: Matthew Benson, Chris St. Laurent, Nicole Noel, Lance Doucette, Stephanie Hoang, Geoff Casey, and Dr. Ian MacDonald.

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March/April 2018 Lab Business

Lab PROFILE of tests to gauge the clinical effect of the vector.

The trial concluded in June 2017 and the team is now pouring over the results.

“This disorder is slow and progressive, so we’re going to see these patients every year and monitor them to find out if the treatment has had an effect,” explains Dr. Ian MacDonald, team leader and Professor Emeritus at the University’s Department of Ophthalmology and Visual Sciences.

The MacDonald Lab includes three research bays, a tissue cultural room and a translational research base. Though much of the equipment tends to be fairly basic, such as thermocyclers for doing PCRs, the lab boasts a fairly extensive cell culture area for viral vector production and one piece of equipment that hadn’t been available in western Canada.

“The Microperimeter is a magnificent machine that would’ve been unavailable to us if we hadn’t undertaken this trial,” says MacDonald. The equipment, funded by the Canadian Foundation for Innovation and Alberta Advanced Education, analyzes the central visual field of the retina by tracking the eye, projecting targets onto the retina at specific points that can be periodically remeasured to determine change. Other organizations that have provided grants to the MacDonald Lab include The Canadian Institutes of Health Research (CIHR), Alberta Innovates, the Foundation Fighting Blindness and the Choroideremia Research Foundation, Canada Inc. MacDonald first began focusing on choroideremia in 1983 while working in Ottawa.

Patients and their families in the eastern Ontario region asked if he knew of anyone who was researching the disease. When he found no one was, he began a journey from gene mapping to gene cloning and then gene therapy.

In 1992, when work took him west to Edmonton, an experience with his neighbour provided additional motivation to continue his research.

“The fence between our properties needed repairs, but I didn’t want to get into that and antagonize the neighbour,” he says. “But he came along with his daughter to see me because she had a problem. Turns out, the daughter was a CHM carrier. Her father was tested and discovered he also had CHM and couldn’t even see the fence.”

CHM affects males significantly more than females because females have two X chromosomes. When one of the female’s chromosomes

The Microperimeter is a magnificent machine that would’ve been unavailable to us if we hadn’t undertaken this trial.

– Dr. Ian MacDonald, team leader and Professor Emeritus, University of Albarta Department of Ophthalmology and Visual Sciences.

carries a faulty copy, the functioning copy will compensate. This means female carriers of CHM typically have mild symptoms and can usually retain good visual function throughout their lives.

MacDonald is still in touch with that neighbour and hopes CHM therapy will safely assist the six patients – all in their early 30s to early 40s – who underwent the trial. But they’re not the only ones eyeing a positive outcome.

Patients have approached MacDonald about researching other disorders that could profit from gene therapy. If the CHM treatment is found to be effective, it could expand the potential to alleviate symptoms of other disorders such as

www.labbusinessmag.com

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Lab PROFILE mental protocol first and then a Health Canada application, whereas Environment and Climate Change Canada has to be involved because these are disabled viruses that are coming into our country.”

Another issue is cost. CHM is only the second disorder in which gene therapy in the eye has been undertaken, the first being Leber’s congenital amaurosis. Spark Therapeutics, a company based in Philadelphia, released Luxturna, an FDAapproved vector to cure this disease for $850,000 (treating both eyes).

“That’s an enormous figure and there will be challenges ahead for the health care system to bring these trials into the clinical domain, but it will happen,” says MacDonald.

Three other centres in the world have conducted similar CHM gene therapy trials and most are accumulating or finishing their trials now. A centre in England selected 30 CHM patients and Germany and Miami performed the therapy on six each.

“When asked whether they would choose to do this again, all of our patients said ‘yes,’” says MacDonald. “They’ve put their eye health on the line as an experiment and have been quite brave about it. We hope there will be some positive benefit for them.” LB

Stargardt’s disease, Usher syndrome and achromatopsia.

“Gene therapy for some of these disorders has already occurred in animals and is now being undertaken in humans,” says MacDonald. “We don’t know exactly what the outcomes will be, but we perceive it will be favourable and maybe one day, gene therapies will be used for both rare and common conditions.”

As for a timeline of when they can deem the therapy a success, Chris St. Laurent, one of the lab’s research technicians, explains the top priority is to ensure the vector’s safety and that it will not cause further damage to the patients.

“This was a safety trial to see whether patients could tolerate the vector and if it could halt further progression in the degeneration of the disease over the next five or 10 years,” says St. Laurent. “That makes it difficult to tell if it’s working immediately, hence the need for such a long period of follow-up in these patients.

“But if there was no trial and these patients didn’t get any intervention, it would be a slow decline into blindness and there’s no way back from that.”

The team at MacDonald Lab continues to work collaboratively with Health Canada and Environment and Climate Change Canada.

“What is very important to Health Canada is that they have an example of a gene therapy trial in the human eye that they can benchmark,” says MacDonald. “They require an experi-

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March/April 2018 Lab Business

If there was no trial and these patients didn’t get any intervention, it would be a slow decline into blindness and there’s no way back from that.

– Dr. Ian MacDonald, team leader and Professor Emeritus, University of alberta Department of Ophthalmology and Visual Sciences.

Application NOTE

Plasma Treatments are

Driving up the Value of Plastic Labware story by

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Jeff Elliott

March/April 2018 Lab Business

Application NOTE

By altering the surface properties of polymer labware, manufacturers are improving the quality of test results while increasing value of products they create

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ach year, billions of multi-well plates, pipettes, bottles, flasks, vials, Eppendorf tubes, culture plates, and other polymer labware items are manufactured for use in research, drug discovery and diagnostic testing.

Although many are simple, inexpensive consumables, an increasing percentage are now being surface-treated using gas plasma or have functional coatings specifically designed to improve the quality of research and increase the sophistication of diagnostics.

Among the goals of surface modification are improved adhesion and/or proliferation of antibodies, proteins, cells and tissue; as well as improved signal-to-noise ratio so testing is more precise with less target material or markers required.

For some labware manufacturers, altering the properties of these devices can also make sense from a business perspective. In a market dominated by several large labware manufacturers, more specialized offerings can create a competitive edge and drive up the value of each consumable. For those creating next generation medical diagnostic devices, coated or plasma treated labware optimized for the testing can improve the quality, specificity and efficacy of the results as well.

“With polystyrene or polypropylene labware, if you can add a functional coating or use plasma to alter the surface properties you can turn a $2 item into a $50 item,” says Mic Barden of PVA TePla America, a leading system engineering firm that designs plasma systems for surface activation, functionalization, coating, ultra-fine cleaning and etching.

Plasma treatment To be sure, some surface modification of plastic labware using plasma is already pursued by the largest labware manufacturers.

Plasma is a state of matter, like a solid, liquid, or gas. When enough energy is added to a gas it becomes ionized into a plasma state. The collective properties of these active ingredients can be controlled to clean, activate, chemically graft and deposit a wide range of chemistries.

Most of the applications of plasma for plastic labware can be categorized as “simple” treatments, such as O2 or Argon for cleaning the substrate at the molecular level. The use of plasma is also well established for surface conditioning to make polymers more hydrophobic (water repellent) or hydrophilic (affinity to water).

However, in vitro diagnostic substrates may require more selective chemistries for the selective adhesion promotion and conjugation of bio active molecules.

This can be achieved by providing particular chemical functionality at the surface, allowing covalent coupling of biochemical species to occur. Amino, carboxylic, hydroxyl and epoxy functionalities are important examples of the chemistries that are readily obtainable using a gas plasma surface treatment.

Multi-well plates Multi-well, or microtiter, plates are a standard tool in analytical research and clinical diagnostic testing laboratories.

Most plates come with 96, 384 or 1536 sample “wells” that function as small test tubes.

The most common material used to manufacture microtiter plates is polystyrene, because it is biologically inert, has excellent optical clarity and is tough enough to withstand daily use. Most disposable cell culture dishes and plates are also made of polystyrene Other polymers such as polypropylene and polycarbonate are also used for applications that must withstand a broad range of temperatures such as for polymerase chain reaction (PCR) for DNA amplification.

Untreated synthetic polymers, however, are extremely hydrophobic and so provide inadequate binding sites for cells to anchor effectively to their surfaces.

To improve biomolecule attachment, survivability and proliferation, they must be surface modified using plasma to become more hydrophilic. Microtiter plates, for example, can be modified with hydroxyl, carboxyl or amine groups to render them hydrophilic (or wettable) and to introduce a negative or positive charge.

Treating the surface in this manner has many benefits, including improved analyte wetting of wells, greater prolif-

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Application NOTE

Most of the applications of plasma for plastic labware can be categorized as “simple” treatments, such as O2 or Argon for cleaning the substrate at the molecular level.

tially unavailable for bonding with antigens.

“With most uncoated polymer surfaces, you can’t control how the Y-shaped “capture” antibodies are oriented,” says Barden. “However, a functional coating can be used to favour the proper upward orientation so the entire surface is available for the assay.

In this way, we can improve the signalto-noise and dynamic range of an assay.” eration of cells without clumping, reduced amount of serum, urine or reagents required for testing and lower risk of overflow and cross-well contamination.

Improved antibody adhesion for bio assays A common usage for microtiter plates is for bio assays such as the enzyme-linked immunosorbent assay (ELISA) used broadly for diagnostic testing. ELISA is used to detect the presences of a substance, usually an antigen, in a liquid sample.

Performing an ELISA involves at least one antibody with specificity for a particular antigen. The sample with an unknown amount of antigen is immobilized on a microtiter plate via adsorption to the surface or via capture by another antibody specific to the same antigen.

After the antigen is immobilized, the detection antibody is covalently linked to an enzyme or can be detected by a secondary antibody that is linked to an enzyme through bioconjugation.

In a final step, the plate is developed by adding an enzymatic substrate to produce a visible signal, which indicates the quantity of antigen in the sample.

To improve the bond and function of the antibody, plasma coatings can be applied to orient the Y-shaped IgG proteins utilized in the majority of these types of tests.

Failure to do so can mean some antibodies face down or deform and become essen-

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Applying an amine coating is a popular solution.

“Amine coatings are commonly used because they have a middle surface energy, with water contact angles of approximately 60 degrees,” says Barden.

“So the coating is hydrophilic enough that the liquid disperses well and hydrophobic enough to facilitate bonding of the material.”

Other alternatives including putting down a linker molecule such as an epoxide or carboxylic acid; or applying a quartz-like surface using plasma enhanced chemical vapour deposition.

According to Barden, all these approaches provide a similar surface energy, but have functional differences that may be important, depending on the application.

Application NOTE According to Barden, plasma treatment is common enough that leading equipment providers are able to modify existing, mature tools and technology, complete with fixturing, to deliver what are essentially drop-in solutions.

crystalline and highly flexible (180o ASTM D522) coating.

Markets for this barrier coating include drug discovery, drug delivery, biological storage, stem cell, and IVF culture wear. In addition to the barrier properties of this coating, SiO2 is also chemically resistant to solvents making it ideal for use in the analytical wear.

Pipette Tips Pipettes are another common laboratory tool. Often constructed of high-density polyethylene or polypropylene that tends to be hydrophobic, pipettes can still have difficulties with liquids sticking to the surface – particularly on, or around, the tip.

To address this issue, some pipette manufacturers add fluorinated polymers within the polypropylene during the injection molding process. However, there can still be issues, such as phase separation or leaching.

To ensure pipette tips “sheet” off any aqueous solution more effectively, companies like PVA TePla can utilize nanotechnology to create a superhydrophobic surface.

One such technique involves etching the surface to roughen it such that air, nitrogen, oxygen and other gases are trapped in the recesses, allowing the liquid to float on the top in a “lotus effect.”

Another method involves applying a more hydrophobic coating on the inside and out of the pipette tip.

PVA has already designed special trays and fixtures capable of treating entire racks for 96 and 384 well microtiter plates. The process utilizes “pulses” of plasma that activate a specific monomer, causing it to diffuse and polymerize within the pipette tip.

R&D assistance According to Barden, plasma treatment is common enough that leading equipment providers are able to modify existing, mature tools and technology, complete with fixturing, to deliver what are essentially drop-in solutions.

Some providers even provide access to on-site research and development equipment as well as engineering expertise. PVA TePla, for example, often invites labware manufacturers to visit its lab in Corona, CA. To run parts and conduct experiments on in-house equipment, with full customer involvement.

It is during these technical customer/supplier meetings that many of the best experimental matrices and ideas are produced.

“The elegance of these [plasma treatment] solutions is that they leverage existing technology and know-how, as opposed to creating something that is completely new,” says Barden.

“Access to this knowledge base facilitates new entrants into the market.” LB

Minimizing leaching Since plastic labware is susceptible to leaching from plasticizers, stabilizers and polymerization residues, plasma is sometimes also used to coat the inside of the containers with a quartz-like barrier material. These flexible quartz-like coatings are polymerized onto the plastic by plasma enhanced chemical vapour deposition.

The resulting coating can be very thin (100-500nm), highly conformal, non-

Jeff Elliott is a Torrance, Calif.-based technical writer. He has researched and written about industrial technologies and issues for the past 20 years.

Surface modification of labware for cell, tissue cultures The enormous growth in studies of whole live cells has led to an entirely new range of microplate products which are cell and tissue culture treated for this work.

According to Barden, the issues of adhesion that apply to proteins used for ELISA can also apply to cells and tissue cultures.

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Lab WARE Tri-Clamp Offers Secure Connection

The latest innovation from Watson-Marlow Fluid Technology Group brand BioPure is the Q Clamp, a new sanitary tri-clamp for single-use fluid path applications.

Capable of providing a secure connection in less than 12 seconds, this patented singlehanded and tool-free sanitary tri-clamp is qualified for use in the pharmaceutical and biotechnology industry. A lightweight sanitary clamp is supported by an industryleading validation pack for non wetted components. The BioPure Q-Clamp does not require any tools for application or removal. Importantly, a double-sided ratchet ensures the clamp remains shut in the unlikely event that one side should fail. www.wmftg.com

Data Analytic Software Solutions Find Hidden Gems

SIMCA has updated its Umetrics Suite to help manufacturing industries make better business decisions and optimize process control. SIMCA, an established advanced data analytics and visualization program as part of the company’s proven Umetrics Suite, makes it possible to combine and analyze data from all sources to isolate, understand and act on the hidden gems that hold the secret to better decision-making and greater business success. The realtime monitoring and prediction system, SIMCA-online, constantly monitors processes to provide a continuous snapshot of the users’ operations. It identifies when set parameters change, enables remedial action to be taken before production is affected and ensures product quality consistency. www.umetrics.com

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Operators Realize High Sensitivity

Filters Reduce Risk of Bioreactor Contamination

MilliporeSigma introduced Viresolve Barrier capsule filters to reduce the risk of bioreactor contamination. Viresolve Barrier filters are designed to remove viruses, mycoplasma and bacteria from cell culture media. Scalable from bench through commercial manufacturing, Viresolve Barrier filters offer high flux and high capacity virus filtration to ensure productivity, without impacting cell culture performance or protein quality attributes.

Available now in sizes up to one sq. m., including scalable micro filters for process development, these capsule filters are easy to implement directly in front of the bioreactor or into a media storage container. www.emdmillipore.com

Shimadzu Scientific Instruments (SSI) recently announced the release of the Nexera Mikros Microflow Liquid Chromatography Mass Spectrometry System. Covering the complete range from microflow to semi-microflow, this system allows operators to realize high sensitivity, yet with the reliability and ruggedness of HPLC. Options include a direct injection system for sample volume-limited analyses for highly sensitive micro LCMS analysis. A trap and elute system is available for analyzes with larger injection volumes. A make-up flow system includes the addition of a post-column make-up flow pump, allowing mobile phase modifiers to be added to ensure the highest ionization efficiency in the MS source. www.ssi.shimadzu.com

Lab WARE Temperature Accuracy and Stability Targetted

Torrey Pines Scientific, Inc. announced its new EchoTherm HPLC Column Oven Model CO20. The temperature range is from room temperature to 90 C readable and settable to 0.1 C. The PID temperature control software regulates temperatures to ±0.1 C. Temperature accuracy and stability are ±0.1 C with a stable temperature indicator lamp on the front panel that lights when the target temperature is stable to within ±0.2 C. The unit operates from 12 volts DC and comes with a benchtop universal power supply for use anywhere in the world. The model is UL, CSA, and CE-compliant. www.torreypinesscientific.com

Ultrahigh Resolution Imaging Combines with Advanced Analytics

ZEISS introduced its new field emission scanning electron microscope, (FE-SEM) ZEISS GeminiSEM 450. The instrument combines ultrahigh resolution imaging with the capability to perform advanced analytics while maintaining flexibility and ease-of-use. Users will benefit from high resolution, surface sensitive imaging and an optical system that ideally supports them in obtaining the best analytical results. In addition, ZEISS GeminiSEM 450 has been designed to cater to a broad variety of sample types from classic conductive metals to beam sensitive polymers.

This makes ZEISS GeminiSEM 450 the ideal platform for the highest demands in imaging and analytical performance. www.zeiss.com

Machine Reduces Risk of Costly Filling Errors

New from Watson-Marlow Fluid Technology group is the Flexicon PF7 peristaltic tabletop aseptic liquid filling machine optimized for operation in GMP-regulated industries such as biotechnology, pharmaceutical and diagnostics. The PF7 adds a more intuitive, validation-friendly operator interface to reduce the risk of costly filling errors. Control could not be easier. Users can adapt filling parameters to achieve optimum accuracy for their application needs, up to 200 user programmable “recipes” can be stored and password protected for future use. A simple and powerful user interface aids operation while working in gloves or behind glass within a RABS or LAF unit. www.wmftg.com

ZEISS Delivers 120 Nanometer Resolutions

In the past, researchers had to acquire a stack of z-slices and subsequently deconvolve to get optical sections thinner than one AU and enhance lateral resolution. Temporal resolution was thus limited, and a prolonged light exposure of the sample was inevitable.

Scientists can now use the new 2D Super-resolution mode to overcome this problem and perform gentle live cell imaging experiments. They benefit from very low light exposure, highly resolved structural information and excellent signal-to-noise ratio. Researchers can process both existing and new ZEISS Airyscan data with the new 2D Superresolution mode. www.zeiss.com

LIST OF ADVERTISERS & WEBSITES Metrohm

Page 2 .............................................. www.blog.metrohm.com

Georgia Department of Economic Development

Page 4 ............................ www.georgia.org/FoodProcessing

Canadian Food Business

Page 9 .............................. www.canadianfoodbusiness.com

Hanna

Page 15 .................................................... www.hannainst.com

Caledon Laboratory Chemicals

Page 23 ................................................ www.caledonlabs.com

VWR

Page 24 ............................................. www.vwr.com/antibody

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Moments in TIME

The Good Doctor In 1961, Dr. Charles Scriver founded the De Belle Laboratory for biochemical genetics at the Montreal Children’s Hospital, which has contributed much to the study of inborn errors of metabolism and genetic disease. Born in Montreal in 1930, Scriver’s work on rickets led to the development of a treatment for the genetic metabolic disease and a preventive nutrition program in Montreal, as well as thalassanemia and Tay-Sachs screening programs.

In the 1960s, he improved the lives of children in Quebec suffering from rickets when he discovered that a vitamin D deficiency was the cause of the disease and its accompanying bone deformation.

Scriver went one step further and convinced the federal government to require that milk producers add vitamin D to milk. He was inducted into the Canadian Medical Hall of Fame in 2001. LB

Reference: https://ville.montreal.qc.ca/ordre/en/charles-r-scriver

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Celebrating In 1971, Doug Brock recognized the need for manufactured high purity solvents. He started Caledon Labs with just a few products, and the goal of becoming a leader in the laboratory chemical industry.

Facility: We have grown from an 800 square foot building to a 20,000 square foot manufacturing facility.

Products: We now provide a wide selection of chemicals, as well as offering custom manufacturing capabilities.

People: Our most valuable asset many employees have been with us for over 25 years.

Quality: A superior standard of quality that our customers have come to expect.

At Caledon, we have built our success on our basic values of Trust, Quality, Service and Delivery. We invite you to experience the Caledon Difference, and allow us to be part of your success story.

Trust. Quality. Service. Delivery.

Caledon Laboratories 40 Armstrong Avenue Georgetown, ON L7G 4R9 Canada T 877.225.3366 E service@caledonlabs.com

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