Scaling Biotech in Minnesota

The Biotechnology Resource Center at 27

Heat, Pressure, Enzymes

Every visit to the BioTechnology Institute features a tour of the Biotechnology Resource Center (BRC), the institute’s 4700 square foot R&D and contract services center. The BRC has grown from humble beginnings in the basement of the Gortner Lab into a state-of-the-art pilot plant performing fermentation process development, recombinant protein expression and downstream processing for clients within the University and beyond.

Celebrating its 27th year in 2013, the Biotechnology Resource Center (BRC) continues to support research at the University while serving as a resource for Minnesota’s biotech industry. Up to 80% of its business comes from Life Science companies ranging from one-person start-ups to some of the biggest names in the biotechnology business.

Income from fermentation services for outside companies helps the BRC fulfill its mission of providing services to the University community at cost, including new equipment and services like a French Press Extruder a Golan press, a Microfluidizer and 550L fermentation tank. Expertise provided by BRC fermentation manager Fred Schendel and his team help scientists involved in basic research develop methodologies for producing molecules at pilot scale—often producing batches of biological compounds at near commercial scale.

Strong connection to industry

Beyond the allure of the centrifuges and stainless steel, visitors quickly learn that the BRC serves an important role in supporting Minnesota’s growing Biotechnology and Life Science economy. And the BRC remains an important standard bearer for the University beyond the Midwest.  In fact, The University of Minnesota alumni network is a primary driver of business for the BRC forming a national and international network from as far away as Uruguay.

Recently the BRC has played an important role in helping companies gear up for clinical trials for chemotherapy compounds and treatments for celiac disease.

From local companies like IGF Oncology, whose founder Hugh McTavish received his PhD from the University of Minnesota to Alvine Pharmaceuticals, a California based company with strong ties to Stanford University, the BRC is able to advance the projects of its partners while saving them capital equipment costs.

IGF Oncology

Like many of the companies served by the BRC, IGF Oncology has a direct connection to the BioTechnology Institute and the BRC. The company’s founder, Hugh McTavish is a PhD biochemist and patent attorney who received his PhD from the University of Minnesota in 1992 and co-authored a 2002 study with BTI’s Mike Sadowsky and Larry Wackett, which identified an enzyme which initiates the metabolism of the herbicide atrazine, a source of ground-water contamination from agricultural run-off.

McTavish is also a cancer survivor and his company seeks to increase the efficacy of existing chemotherapy by creating drug conjugates, which help target cancers cells while reducing damage
to healthy cells. If successful, IGF’s product holds the promise of lower dosages and fewer side effects than current chemotherapy compounds. Animal studies have shown the IGF conjugate to be effective at dosages 6 times lower than the chemotherapy drug methotrexate alone.

When IGF was planning Phase I clinical trials they turned to the BRC to produce a sufficient amount of the conjugate to conduct a trial on 20-30 patients. Using the strains provided by IGF,
the BRC scaled-up a fermentation and purification process from bench scale to production levels necessary first for animal studies and then to 250 liter scale to produce the material needed for a phase I trial.

Alvine

Founded on technology licensed from Stanford University, Alvine Pharmaceuticals develops therapeutic compounds targeting autoimmune/inflammatory diseases. The company’s current focus is celiac disease, a chronic condition induced by the protein gluten found in wheat, and related grain species like rye, and barley. Gluten contains high concentrations of two amino acids, proline and glutamine, which are not broken down efficiently in the stomach and small intestine. Peptides absorbed through the small intestine cause an autoimmune reaction in individuals susceptible to the disease. Alvine’s flagship product, ALV003 contains protease enzymes engineered to digest gluten. In 2009, when Alvine entered Phase II clinical trials, it turned to the BRC to produce cell paste with the recombinant protein expressed. Using a working cell bank supplied by Alvine, the BRC grew the cells at pilot scale to Alvine’s specification in several 500 Liter reactor runs. The cell paste was then sent to Alvine for further purification.

Building on the success of the initial engagement, Alvine returned to the BRC in 2010 for assistance developing an economical process for scaling up to commercial production at Alvine’s own facility. The engagement, completed in 2012, underscores the value the BRC provides in helping bring novel products from early testing phase to commercialization and large-scale distribution.

“The BRC is not just a business running within the University,” according to BRC Director Tim Tripp. “Outreach to industry is essential to our mission. Another key component is our ability to provide training in processes not available elsewhere at the University.”

The Short-Course

Coaxing an organism to produce a novel compound in a petri dish or shake flask doesn’t guarantee success at a commercial scale. A host of variables including feedstock, temperature, and acidity levels all contribute to the successful transition from lab bench to commercial scale. Through its short course, the BRC provides training in bench-top fermentation processed to students within the University and companies seeking to improve in-house fermentation capabilities.

The course is taught by Fred Schendel, a PhD level fermentation biochemist with over 10 years of experience in industry, along with BRC  Director Tim Tripp, and a rotating group of faculty members from the BioTechnology Institute. The course is open to students within the University and industry.

In addition to lectures and lab visits, students work in teams to design and complete a fermentation using one of the BRC’s  five liter bench scale fermenters. As Tripp points out,
“The BRC is the only place on campus where students can find structured, hands-on fermentation training, a valuable skill in Minnesota’s growing Life Science economy.”

A Biotech Catalyst

In addition to its weeklong short course on bench-scale fermentation and services offered through the pilot plant, the BRC is also home to a growing number biotech companies through Lab Use Agreements offering affordable, small-scale lab space and direct access to experts within University’s academic community and the BRC.

Butrolix

Founded in 2008 by Don Mattsson, PhD, a former UMN student, and one of the first three graduates of the BioTechnology Institute’s Masters in Microbial Engineering (MicE), Butrolix (www.butrolix.com) was one of the first companies to rent space from the BRC.

While completing his doctoral and postdoctoral training at the University of Minnesota, Mattsson discovered the molecular components used by butanol-producing bacteria to regulate biobutanol formation. In 2008, after spending time in industry, Mattsson and his wife, Attorney Lisa Mattsson, formed Butrolix with the goal of developing a patentable process for biobutanol production from low-cost sugar feedstocks. In fact, Butrolix was the first company to patent the use of quorum sensing peptides to synchronize bacterial populations and increase the speed of butanol production.

Dr. Mattsson sites the resources offered by the BRC as a key factor in getting Butrolix off the ground. After researching facilities closer to his native Duluth, Mattsson choose the BRC because of the access it provided to the BRC’s fermentation facility and the expertise of the BRC staff and faculty.

When Butrolix sought a Phase I SBIR award from the National Science Foundation (NSF), Mattsson was able to draw on BTI director Mike Sadowsky, who provided a supporting letter for the NSF, while the BRC was listed as subcontractor.

CTE Global

CTE Global Inc. is a manufacturer of industrial enzymes with production facilities in the U.K., China, and Brazil and 150 enzymatic preparations used in over 15 industries in 48 countries. Its fermentation products are produced from plant material, microbes, and fungi including an Aspergillus derived enzyme that facilitates the fermentation of ethanol, butanol, organic acids and other specialty chemicals. Preparation used in ethanol production help producers reduce hazardous chemical inputs and use costly feedstocks more effectively, improving the bottom line while reducing energy use and improving air and water quality.

As the company sought to extend its market share in they increasingly competitive ethanol industry, they began looking for a Midwest facility to focus on outreach, quality control, and product development.

Proximity to clients played a major role in CTE’s decision, but the BRC provided a number of other advantages, which sealed the deal, including affordable, move-in ready lab space with critical infrastructure, logistical and administrative support from Director Tim Tripp, and the availability of contract services through the fermentation pilot plant.

Add to this access to the expertise within the University and a wide variety of sample feedstocks available at the University’s St. Paul agricultural campus, and the BRC was a perfect fit for CTE’s first U.S. Quality Assurance facility.

In the U.S., one of the primary goals is to protect against contamination in transit. Staffed by Dr. Sandra Lobo, a biochemist with strong Minnesota ties, CTE has stocked its St. Paul facility with state-of-the-art equipment to help ensure its enzymes preparations are protected against contamination in transit. Beyond quality control and assurance, Lobo plans to use the facility to develop new enzymatic formulations to enhance biocatalysis for the wide variety of cellulosic feedstocks used in ethanol production.

Lipodome

Lipodome, the most recent company to enter a lab use agreement with the BRC, was founded in 2001 by Tarun and Napur Ghosh. The company is currently focused on developing and marketing its product line, used to facilitate the study of proteins and lipids in the development of therapeutics for inflammatory diseases and cancer. In addition to its platform technology for solubilizing detergent sensitive membrane bound proteins and enzymes, Lipodome also markets products which allow the retention of protein functionality in a stable, non-detergent buffer medium. LLS KitsTM  allow for the separation and quantitative analysis of  lipids 10-20 times the rate of conventional assays.

Going Global

BTI Annouces new 5-year academic exchange agreement with Osaka University’s Institute of Scientific and Industrial Research

Building on the success of its academic exchange programs with the Nara Advanced Institute of Science and Technology (Nara, Japan), BTI recently announced a new 5-year Academic Exchange Agreement with Osaka University’s Institute of Scientific and Industrial Research (ISIR). The exchange program will include collaborative research, symposiums, and the exchange of researchers, including undergraduate and graduate students.

The ISIR has a strong international focus with an emphasis on basic and applied research related to energy, the environment, and the biological and molecular sciences. Interdisciplinary in nature, the institute includes researchers from the Departments of Microbiology, Material Sciences, Nanotechnology and Information Sciences and is part of collaboration network between Japanese national universities including the Institute for Materials Chemistry and Engineering at Kyushu University and the Chemical Resources Laboratory at the Tokyo Institute of Technology.

According  to BTI Director Mike Sadowsky “ISIR is a good partner for academic exchange for BTI. Both share a cross-disciplinary approach to basic and applied research.”

The collaboration began this summer when BTI hosted the first group of five senior researchers from ISIR in a 2-day program featuring formal presentations by BTI and ISIR. The visit also included series of breakout sessions were members of ISIR met individually with BTI members with similar research interests. According to Sadowsky, who launched the program after an exploratory visit to ISIR in 2012, the breakout sessions are extremely important in forging personal bonds between international researchers and in exploring potential areas for collaboration.

The symposium included presentations by BTI members Daniel Bond,
Jeff Gralnick, Igor Libourel, Romas Kazlauskas, Yiannis Kaznessis, Dan Knights, Mike Sadowsky, and Michael Travisano. ISIR presentors included:

Yashushi Yagi (ISIR director, Department of Intelligent Media), who gave and overview of ISIR’s mission and research and discussed his research Yagi focuses on computer vision and pattern recognition and advances in computer imaging using parallel high-frequency illumination.

Nobuo Kato (Department of Organic Fine Chemicals) spoke about his lab’s efforts to stabilize protein-protein interactions with the possibly of developing drug targeting technologies.

Takeharu Nagai (Department of Biomolecular Science and Engineering) presented his research on Nano-lanterns and luminescence indicators for bioimaging to help solve the problem of autofluorescence in the imaging of plant microbe interaction.

Kunihiko Nishino (Laboratory of Microbiology & Infectious Diseases) who presented work on multi-drug efflux pumps in Salmonella enterica and their relevance to antibiotic resistance.

Kazuhiko Nakatani (Department of Regulatory Bioorganic Chemistry) presented his work on base pair mismatching and the introduction of single nucleotide polymorphisms as a way to develop desirable phenotypes in plants.

BTI Assistant Director Tim Tripp commented, “The ISIR-BTI symposium demonstrated that the language of Life Sciences transcends national and regional culture. Uncovering biotechnology solutions to global problems will require the kind of international collaboration begun by BTI and ISIR.”

The 3rd Dimension

The 3rd Dimension

BTI Researchers test 3-D printing technology to scale up—and down

Since it first appeared on the market in 1984, 3-D printing technology, also known as stereolithography, has been used to create everything from robotic aircraft to artificial limbs. The technology caught the attention of the media this summer, when reports surfaced that a Canadian man fired 14 shots from a rifle manufactured on a 3-D printer using a design downloaded from the Internet.

The cost of the technology is becoming more affordable—desktop units now range from $250-$2500—and the printers are finding their way into artist studios, research labs, and in some cities, the local Fedex/Kinkos copy shop.

In 2012, BTI members Brett Barney (BTI/Department of Bioproducts and Biosystems Engineering) and Igor Libourel (BTI/Department of Plant Biology) approached BTI director Mike Sadowsky for funds to purchase a MakerBot Replicator 2—a low-cost 3-D printer about the size of bread box. Both saw the potential to advance research and training goals but each had a novel approach to experimenting with the technology.

Barney immediately saw the potential of the technology in the classroom and has created brightly colored, hand-painted models to help explain cell metabolism and metabolic pathways to his students.

Beginning with three dimensional images of proteins from the Protein Data Bank, a repository of structural images for large biological molecules, he uses a variety of

3-D rendering tools to refine his models and build the scaffolding required to support the structure as the printer lays down layer upon layer of ASB thermoplastic, similar that found in Lego® building blocks. After cutting away the scaffolding and the excess material, the molecular models are painted, polished and ready for display.

The initial models took up to 15 hours to print, with several additional hours of detailed work for clean up and finish. With practice, he was able to reduce the printing and clean up cycle to a couple of hours. Once the models are complete, Barney posts the plans, with photographs and annotations, on Makerbot’s open-source repository called Thingiverse. Published under the name MoleculeMaker, Barney’s model of the FeMo Cofactor is available for download by members of the online community.

Barney hopes to print a full model of a photosynthetic reaction center, but also builds partial models to highlight unique characteristics of the molecules he studies. In fact, his lab has used the models to help predict sites for mutagenesis studies in wax ester synthases—enzymes important in the effort to produce biodiesel from a complex biomass such as cellulose. Barney recently published the
results in the Journal of Applied and Environmental Microbiology.

For more about Barney’s 3-D Models see: http://barneybioproductslab.cfans.umn.edu/3d-printing-and-molecular-models/ 


Rapid prototyping and development of miniature bioflow reactors

With an eye toward understanding environmental changes brought about by global warming, the Libourel Lab studies metabolic features of Ostreococcus, a picoalgae genus common in the world’s ocean. Investigating the relationship between metabolic adaptation and climate models, the lab relies on bioflow reactors to manipulate and monitor the organism’s response to evolutionary pressure.

After modifying bench scale reactors with customized hardware and software, Libourel and his students realized they could achieve the same results, using fewer resources, by scaling down. But each bioreactor requires a custom enclosure, which can cost as much as $500 to produce, increasing cost and slowing development. Using the Replicator 3-D printer, Libourel hopes to construct and modify the enclosures which house the circuits, pumps, and fans required to run the reactors. In addition to the cost saving, the rapid prototyping and development will allow the lab focus on what’s happening inside the dish instead of the box.

BTI’s Wei-Shou HU spearheads new consortium to speed drug development from Chinese Hamster Ovary cell lines

Distinguished McNight Professor Wei-Shou Hu (BTI/CEMS) is leading a new Consortium for Chinese Hamster Ovary Cell Systems Biotechnology formed under the auspices of the Society of Biological Engineering. This new consortium of seven pharmaceutical companies from USA, Europe and Japan is built upon work of the proceedings of the Consortium of Chinese Hamster Ovary Cell Genomics from 2006-2010, which completed sequencing of the Chinese hamster genome and developed platforms for genome scale analysis.

The Chinese Hamster Ovary (CHO) cell line is used in the production of over 70% of all biologics, valued at over $40 billion per annum.  The new effort aims to harness the genomic information and further advance our understanding of cell line production, productivity and product quality and to facilitate member companies entering the post-genomics era of biologics manufacturing.

Gary Muehlbauer named distinguished McKnight Professor

BTI member Gary Muehlbauer, was recently named a Distinguished McKnight Professor. This award is granted to outstanding faculty members who have recently made the transition to full professor status. Muehlbauer is head of the Department of Plant Biology, and is also a part of the Department of Agronomy and Plant Genetics. His research is concentrating on wheat and barley and molecular genetics, as well as integrating genomics resources into barley breeding programs. Highest regards to Gary Muehlbauer.

Kechun Zhang joins the BioTechnology Institute

Kechun Zhang

BTI/Chemical Engineering and Materials Science BTI welcomes Kechun Zhang, Assistant Professor in the Department of Chemical Engineering and Materials Science. Zhang completed his PhD in chemistry at the California Institute of Technology and his postdoctorate fellowship at the University of California, Los Angeles. Zhang’s research focuses on the sustainable production of biofuels, chemicals, and biologics through engineered processes including protein evolution, metabolic flux, and the design of artificial metabolic pathways. In 2013 Zhang was the recipient of the 3M Non-tenured Faculty Award and the American Heart Association National Scientist Development Grant Award.