Partnerships Strengthen Finland Biotech
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Genetic Engineering News
Apr 15 2009 (Vol. 29, No. 
Partnerships Strengthen Finland Biotech
Country’s Network of Collaborators Is Far-Reaching and Ultimately Critical to Success
Susan Aldridge, Ph.D.
Finland, particularly the Oulu region, is focused on developing the connections between the life sciences, nanotechnology, and information technology. Oulu Bioforum, Oulu Innovation, and other centers in Finland, supported by partners in the German cities of Halle and Leverkeusen, recently hosted “Bio Meets Nano and IT” to promote interdisciplinary and business collaboration between those involved in these three areas.
High-throughput approaches are a special interest in the Oulu region. Hans Söderlund, Ph.D., VTT Technical Research Center described the development of a high-throughput multiplexed system for analysis of transcriptional signatures that is based on advanced microfluidic technology.
“To measure total gene expression by conventional means is cumbersome and expensive,” he said. There are gene chips that can measure 40,000 genes and, at the other extreme, those that look at just a few. “We are interested in the space between 10 and 300 genes, looking at a large number of samples with high throughput.”
Transcript analysis with affinity capture (TRAC), a VTT technology, has been spun off to PlexPress TRAC involves exposure of RNA through cell lysis and then the addition of a capture probe that binds to streptavidin-coated magnetic beads. This way, data on 2,880 transcriptional levels (96 samples in a 30-plex experiment) is available in about four hours, Dr. Söderlund reported. Applications include cancer cells exposed to drug candidates, RNAi knockdown assays, and cell-cycle monitoring.
In a collaboration with the University of California, Berkeley, the company has transferred TRAC onto nanovolume chips for online analysis and is also trying to transfer the technology onto a printed format, reflecting VTT’s interest in getting wet chemistry onto chips to make low-cost biosensors. “We could, therefore, go up to high-throughput transcriptional profiling,” added Dr. Söderlund.
Olli-Pekka Kallioniemi, Ph.D., of the Institute for Molecular Medicine discussed the Institute’s medical systems biology approach, with the development of a high-throughput siRNA screen of cancer cells in 384-well plates. Scientists at the Institute are miniaturizing this to a functional cellular microarray where they will print siRNA from 20×384 well plates to see how the cells respond to the siRNAs.
“We can do the entire genome of siRNAs on a single microtiter-sized plate,” he said. The cells adhere to the spot and are transfected by the siRNA. The work has been applied to a prostate cancer cell line in a druggable genome screen. “This is pushing the limit of anything available today. We are now beginning to approach reasonable accuracy and reproducibility.”
In a collaboration with the University of Turku the team is looking at what genes are necessary for integrin signaling when cells invade. Live cell imaging of activated integrin siRNA hits in P13 cells can reveal what happens in cell morphology when this gene is knocked down.
“We use this to identify critical determinants of cell movements,” Dr. Kallioniemi said. “There are still lots of challenges, including the printing requirements for 40,000 siRNAs per experiment and managing up to 40,000 high-resolution images per experiment.” A further application is to improve the efficiency of hormonal therapies in prostate cancer by looking at the siRNAs that have an effect in androgen-deficient conditions.
Lab-on-a-Chip
Oulu Innovation focuses on economic development in the high technology and high growth business sector in the Oulu region. Lab-on-a-chip developments were discussed by Claude Vauchier of the Lab-on-a chip division at CEA-LETI. Smartdrop is based on electrowetting on dielectric technology, which allows actuation on individual droplets (60 mL to 2 µL), opening up the possibility of allow miniaturization and integration of various technologies like RT-PCR applications.
Meanwhile, Marko Pudas, Ph.D., CTO and cofounder of NanoGalax, described how his company has found a way to measure the adhesion of a single living cell to the extra cellular matrix (ECM) via integrin molecules. GalaxCell™ is based upon a piezo actuator and supporting microelectronics. It detaches the cell from ECM observed under phase contrast microscopy. This is potentially useful in cancer research to monitor the effect of drugs that can change the binding of cancer cells to the ECM. It could also be used in cell therapy as adhesion data is informative about the state of a cell’s differentiation. The work is now being extended to micromanipulation applications to gain further insights into cell properties.
Also in this space is SensApex, a spin-off from the University of Oulu that focuses on precise sensing, measuring, and handling/manipulation. The company’s first product is a remote-controlled and ultracompact micromanipulator. It offers stable manipulation in 3-D with sub-micrometer resolution and up to 18 mm movement range per axis. The system comes equipped with a stand-alone control unit with a location display and step-size adjustment.
Cell Cultivation
Biocatalyst-controlled cultivation of E. coli with EnBase in a 96-well plate Developments in bioprocessing coming out of the University of Oulu led to the founding of BioSilta, which has developed a bacterial cell-cultivation technology based on a fed-batch method used in industrial processes. In EnBase™ any well of an MTP plate becomes a miniature bioreactor with a storage layer of starch-containing gel on the bottom and a liquid media layer containing an enzyme.
The enzyme acts as a pump that controls glucose release from the starch to the culture. This controlled nutrient-feeding technique provides much better conditions for bacterial cell culture, the company says, allowing up to 50 times the cell density and up to 10 times higher levels of recombinant protein compared to standard cultivation methods.
“EnBase allows you to scale down your research knowing that you can readily scale it up again,” explained CEO, Russell Golson. The product is available for a variety of formats: shake flasks, mini-shake flasks, starter tubes, and a range of deep well and standard MTP plates. A 24-well Opti-Set allows customers to try different glucose feeding concentrations alongside different media in a single-plate experiment.
“Cell culture conditions can be optimized in a short space of time,” added Golson. Applications of EnBase include increasing expression levels of recombinant proteins (with the University of Regensburg), achieving higher growth for B. subtilis (with the University of Greifswald), and increasing throughput on the Human Proteome Resource with collaborators at the Royal Institute of Technology in Sweden.
In another cell culture development, Peggy Stock, Ph.D., researcher in the molecular hepatology group at the University of Halle, described CellTech-BioReactor, a new system for biochemical monitoring. The project, funded by the state of Saxony-Anhalt, is being developed by the NEMO network, a consortium of small German companies and research facilities. The CellTech-BioReactor consists of a small disposable bioreactor allowing rapid assessment of the impact of compounds on hepatocytes in 3-D culture on various scaffolds.
A sandwich hybridization assay detects changes in liver enzyme (CYP) gene expression that could be related to compound toxicity. The system, at the prototype stage, is highly specific for CYP type, according to Dr. Stock, who hopes it can help replace animal systems.
Biomolecular Interactions
SensApex’ first product is a remote-controlled and ultracompact micromanipulator. Analysis of biomolecular interactions were also discussed. Markku Kulomaa, Ph.D., of the University of Tampere presented new types of avidins, a chicken-derived protein used widely as a probe in the study of biological macromolecules. Both avidin and its bacterial analogue, streptavidin, have been modified by mutagenesis for improved properties.
For instance, the avidin-biotin technology would benefit from avidins with multiple binding sites, said Dr. Kulomaa.
Temperature and organic solvent stable avidins have been created using an approach that compares amino acid sequences of avidin and similar proteins. A number of novel and multipurpose avidins are now being produced by this approach, some of which (the antidins) utilize ligands other than the well-known biotin.
Novamass, an ADMET specialist company, is now a member of Systems Biology Worldwide, a joint Finland-India venture addressing many aspects of early drug development. Novamass continues to develop its core competence in drug metabolism analysis, with a focus on metabolite identification.
Relationship with India
Connections between Finland and India are growing stronger. One of the Indian companies at the meeting was Avesthagen, whose vision is to bring about the convergence of population genetics, functional foods, and pharma for personalized, predictive, and preventive healthcare. Its leading project is Avestagenome™, which involves biomarker discovery and predictive diagnostics. This involves the 70,000 strong Parsi population with its well-defined genealogy. The group is known for longevity, but also for increased incidence of neurological disease, stroke, heart disease, and certain cancers.
“This is the first population-based study with a global impact using a systems biology approach,” said Manan Bhatt, vp of external relations. The current focus of the genome project is upon Alzheimer’s, Parkinson’s, and breast and prostate cancers, with 20,000 samples to be collected by 2010. Currently, Avesthagen is carrying out genome, transcriptome, proteome, and metabolome analysis on samples collected for the Avestagenome project, and is in talks with Finnish and other collaborators on application of chip technology for designing molecular diagnostics.
Anna Erkkilä, head of the trade center and senior consultant of FinPro India in Mumbai, discussed the healthcare market in India and opportunities for life science companies. FinPro is a networking organization dedicated to internationalizing Finnish companies. She observed that demographic change was leading to a diamond-shaped healthcare market in India, with a big increase in middle-income consumers. “Companies go to India because of the reduced cost, but now stay for the quality and competitiveness,” she said.
The diagnostics market is growing, particularly in the in vitro diagnostics area, the driver being an increase in health awareness among the population, although the challenges differ in rural areas. Imported products are often too expensive, so local production is the key. Needs include diagnostics for TB, HIV, malaria, heart disease, cancer, and diabetes.
In rural areas of India, the lack of a cold chain, long distances to transport materials, and high temperatures, as well as a lack of doctors, provide challenges.
Finland will grow its presence in India through funding available for Finnish-Indian early-stage companies in medical diagnostics from Tekes, the Academy of Finland, and the Department of Biotechnology, Ministry of Science and Technology, India.
(previous article, 3/08):
Oulu Region Lures Biotechnology Firms
Finland Seeks to Build a Biotech Cluster to Complement Established Wireless Industry
Susan Aldridge, Ph.D.
It is not surprising that Oulu, Finland, a global center for wireless technology, has a vision of bringing biotech, information and communications technologies, as well as nanotech together. The Oulu region, however, is also strong in more traditional biotech areas such as bioprocessing, GMP manufacture, and drug discovery and development.
Oulu’s two leading assets are Biocenter Oulu (BCO) and Technopolis. BCO, founded in 1986 within the University of Oulu, was Finland’s first biocenter. It focuses on research in biotech and molecular medicine. BCO was also a driving force in the recent establishment of Biocenter Finland, a network of six Finnish universities.
Technopolis is the largest provider of operating environments for high-tech companies in Europe. The three Technopolis centers in the Oulu region have developed a growing interest in adding the life sciences to their high-tech profiles.
Creating a Strong Biotech Cluster
It is the job of Bioforum Oulu to bring together companies, interest groups, and infrastructure to build a strong biotech cluster, driving forward the regional innovation strategy in the life sciences. Bioforum is a member of the national HealthBio cluster in the Finnish Center of Expertise Program. It covers human genetics, diagnostics, drug discovery and development, gene therapy, pharmacogenomics, and regenerative medicine.
Finland’s bio business sector comprises 120–150 companies with a total turnover of E330 million, of which E185 million is in the Bioforum focus fields, says Tuula Palmén, Ph.D., Bioforum’s program director. The bigger companies include Orion, Thermo Fisher Scientific, GE Healthcare, Bayer Schering Pharma, and PerkinElmer. She explains that Oulu’s strengths in biotech include value-chain thinking and using strong and commercially interesting academic research as a source of innovation.
There are 33 biobusiness companies in the Oulu region. BioSilta, formerly Aurora BioTec, founded by Peter Neubauer, Ph.D., professor of bioprocess engineering at the University of Oulu, is one of the newest.
“A small country like Finland, unlike the U.K. or Germany, must focus,” he asserts. “This is well understood here. The cost of R&D per head may seem large, but the total amount is relatively small. We want to bring this region forward with clusters and platforms for discovery, such as the Bioforum.”
Microbial Production
Dr. Neubauer has a long-standing interest in the manufacture of difficult recombinant proteins such as Wnts and collagen. He is closely associated with the BCO through his work in bioprocessing, emphasizing microbial production. “We can make E. coli better able to make all folded proteins. In the future, we will make the periplasm do the job of the endoplasmic reticulum. Even today, we can do a lot in this area.”
Currently, bioprocess development in E. coli takes a long time and is not straightforward. “In the future, we must integrate computer-based models and parallel approaches together with the competence from cell physiology and bioprocess development,” Dr. Neubauer notes.
He points out that one way around this may be to introduce fed-batch production earlier on in development to save optimization time later. His team has been developing such tools, which led to the founding of BioSilta.
For example, SENBIT® is a modular system involving the use of sensors in shake flasks based on a wireless system that allows it to be used anywhere in a facility. Another tool is a controlled-substrate delivery system, which enables cell cultivation to high density, avoiding the toxic osmotic effects of adding large amounts of glucose substrate. “This is an absolute breakthrough technology,” according to Dr. Neubauer.
He is also developing analytical methods for quantitative mRNA analysis from whole cell extracts, which will help to control and understand the transient and dynamic responses in a bioprocess.
Contract Manufacturing
Medipolis GMP, a privately held CMO, specializes in pilot-scale cGMP microbial fermentation of biologics in E. coli, P. pastoris, and B. subti. The company offers expression system and process development studies for biopharma companies, thereby converting bench-scale to pilot- and commercial-scale production for therapeutic proteins, enzymes, and vaccines.
Completed in 2001, the company’s biopharmaceutical facility comprises 2,300 m2 of space, of which 400 m2 is cleanroom. It meets FDA and EMEA requirements and is also approved by the National Agency for Medicine, Finland. The plant was upgraded for commercial production in 2006. “We want to become an expert group in recombinant protein manufacture within the next five years,” says Ashesh Kumar, Ph.D., director, biopharmaceuticals.
The Medipolis production site comprises two 15 L, one 150 L, and one 750 L fermenters. The firm says that it has manufactured the largest amount of therapeutic recombinant protein in Finland, successfully producing Phase II material for clients in the country and internationally, who partner for their fill-and-finish operations.
Dr. Kumar says there are some interesting new manufacturing opportunities on the horizon, such as biosimilars and new types of insulin. “Our mission is to help our customers bring their products to the market faster and more cost efficiently than other companies by offering reliable partnerships in manufacturing.”
CRO Presence
Novamass is a contract research organization (CRO) specializing in ADME studies and was formed as a spin-off from the University of Oulu in 2000. The firm received funding from TEKES, the national development agency for technology and innovation, to develop in vitro models and analytical methods to investigate lead compounds for Juvantia Pharma and Hormos Medical, its first clients.
“The company emerged from our customers’ needs,” says Jouko Uusitalo, CEO. “We solved problems for them, developed new service products, and offered them a good package of in vitro ADME.”
Novamass’ services focus on late discovery and early development. Its products carry out studies such as those on the basic properties of a compound and in vitro metabolism, with the identification of metabolites being a speciality. “We can also predict whether a customer’s compound will interact with other drugs,” Uusitalo says.
“We create added value to the customer by tailoring our methods to their compounds,” he continues. “If there are problems, they get to know about it sooner.” He explains that most companies, especially those in high-throughput sequencing (HTS), deal mainly in numbers and don’t explain results that can be hard for small biotechs to understand. “Our reports are more like a scientific article. We explain what has to be done next.”
Novamass is the only company in the Nordic region with 45 biotech customers in 11 countries including India and many in Europe like KuDOS Pharmaceuticals and Cyclacel. The firm is on the verge of expansion with new ownership and the purchase of an in vivo business.
Uusitalo says they want to grow organically and through M&As. Novamass is currently negotiating with companies in Finland that are involved in areas like GLP and animal as well as clinical trials organizations. The firm also plans to work on HTS in India. “We will do studies in Europe that need to be close to the customer, and India can be cost effective,” he says.
Drug Discovery
Besides manufacturing and drug development, there is a great deal of interest in drug discovery in Oulu. The collagen research unit (CRU) at the BCO consists of three groups. Studies on collagens and collagen enzymes was instrumental in setting up FibroGen, the recombinant collagen specialist with a focus on biomaterials and fibrotic diseases.
Recently, the CRU, in collaboration with FibroGen, has shifted its attention to anemia and ischemia, according to Johanna Myllyharju, professor of molecular biology. They have been investigating an enzyme known as collagen prolyl 4-hydroxylase (P4H), which is essential for processing collagen into its correct triple helical structure.
More recently, a separate P4H family with another role was discovered. Under normoxic conditions, this newly identified P4H group helps break down a transcription factor called HIF, which otherwise regulates EPO and VEGF and helps the body to survive hypoxic concentrations.
There are many potential therapeutic opportunities for HIF in conditions associated with hypoxia, such as anemia, myocardial infarction, stroke, renal hypoxia, and cytoprotection. The strong background of the CRU in collagen P4Hs has enabled them to look for inhibitors of mechanistically similar HIF P4Hs that would allow HIF to survive and carry out its role. “Essentially, such an inhibitor wou ld help to mimic the state of hypoxia response in tissue,” explains Myllyharju.
Working on Lab-in-Pocket Devices
At the VTT Technical Research Center of Finland, Markku Känsäkoski, senior research scientist in the biomicrosystems section, and his team are developing devices involving printed-intelligence. They predict that these could be used in rapid diagnostics in medical and environmental applications with advantages of low cost and high repeatability.
The instruments involve the use of micromechanical components such as pumps fixed in micrometer-scaled channels. “There is still a big gap between biotech and hardware engineering,” Känsäkoski points out. “We are currently working with model assays in areas such as cardiac markers. The hope is that the research could lead to the lab-in-pocket type of disposable diagnostic, a vision that will involve several VTT groups in printed electronics, microfluidics, optics, and antibody libraries.”
To help build and strengthen the Oulu cluster, Finland recognizes the need to create the best possible networks of financing, market analysts, and customers in the main markets. Additionally, using the best expertise and contacts to make the R&D period shorter is important, Palmén adds.
The country differs from its Scandinavian neighbors as it has less of a home-grown pharma industry. Yet, they all face the same challenges in obtaining financing for early-phase innovation. Finland would like to have more access to funds such as Inveni Capital, a new VC company with offices in Finland and Germany. A recent example of progress in this area is a partnership between the University of Oulu and IP Finland, bridging business and financing networks to innovations.
Palmén recently returned from a trade mission to India, a country seen as an increasingly important partner for Finland. “It is a huge, emerging market with additional features important to us,” she says. Finland’s bio sector also collaborates extensively with North America, Germany, and the U.K. Nationally, the HealthBio cluster is seen as an important partnering organization.
Susan Aldridge, Ph.D., is a freelance science and medical writer specializing in biotechnology, pharmaceuticals, chemistry, medicine, and health. Web: www.aldridgeassociates.co.uk. E-mail: susaldr@aol.com.