On January 1, 2000, Dutch diagnostics executive Tim Kievits gave birth to a millennium ‘baby’: a start-up company, based in Den Bosch, Netherlands, organized to develop and commercialize the flow-through microarray technology that he and colleagues at Dutch diagnostics company Organon Teknika had incubated over the past four years.
The new baby would be named PamGene, not for a woman as some might suppose, but after an acronym for poly-analyte measurement, the original term used by the development team to describe the function of the chip they were developing, which is now called the PamChip.
With the birth of PamGene Kievits left his secure job as director of technology for nucleic acid diagnostics at Organon, a division of Dutch conglomerate Akzo Nobel, where he had been working for 10 years.
But despite the culture of lifetime loyalty to companies that still lingers in Europe, Kievits says he wasn’t afraid to make the leap. “Those who stayed behind, those are the ones who take the real risk,” he said confidently during an interview with BioArray News in Zurich last week. “I am a big supporter of spinoffs and spinouts and getting more experience in your life.”
For Kievits, the risk soon paid off. By May, Dan van der Heyden, a former Organon colleague had joined him to become chief financial officer. And by December, the company had closed on 6.7 million, ($5.9 million) in first-round financing from three investors, GIMV in Belgium, Life Sciences Partners in the Netherlands, and Alta Partners in San Francisco. The Belgian investors combed carefully through the financials and contacts, Kievits said, while Alta partnerJean Deleage, focused on face time, meeting with Kievits and discerning based on personal interviews that he had a solid strategy he could execute.
In March 2001, Olympus Instruments of Japan, with whom Kievits had longstanding contacts, added its vote of confidence by agreeing to manufacture an instrument to read the chips and to market the instruments and chips together.
Nourished with support from east and west, the fledgling company began a rapid growth spurt. Currently, PamGene has 27 employees, and plans to more than double by the end of the year, Kievits said. The company is building a manufacturing facility on site in Den Bosch, and is hiring employees for the manufacturing operation. In the second quarter, Olympus is slated to begin marketing PamGene’s PamChip along with its instruments, so the company has to ramp up its manufacturing capabilities quickly.
Meanwhile, Kievits and his management team have signed a couple of four-year partnerships in their own backyard, including one with Erasmus University Medical Center in Rotterdam to apply the PamChip to osteoporosis and osteoarthritis and another with the Royal Tropical Institute (KIT) in Amsterdam to develop a PamChip that focuses on detection of antibiotic resistance markers. To support these projects, the company received a $3 million grant from the Dutch government in July.
They have also developed a diagnostic prototype of a PamChip that detects drug-resistant strains of Mycobacterium tuberculosis, which they presented at the biochip conference in Zurich last week. The chip has species-specific probes for strains of Mycobacterium tuberculosis and for other Mycobacterium strains. It offers advantages over other diagnostic methods, according to Alan Chan, the company’s vice president of scientific applications, because it is more sensitive than standard direct microscopy, faster than cell culture, and can probe for more species in parallel than current molecular techniques.
‘Tree Trunk’ Microarrays
The PamChip’s core technology is a porous material somewhat similar to a cross-section of a tree trunk, with natural capillaries running vertically through it. The company can manufacture large sheets of this substrate, reducing cost per chip. A non-contact piezo-inkjet printer deposits oligonucleotides in these capillary channels, and the oligos cleave to the sides. Then a drop of target analyte is added to the channel and a microfluidic pump “pulses” the drop so it moves up and down through the capillary.
This rapid diffusion is designed to speed hybridization times, and takes less than an hour — or in the case of the Mycobacterium chip, under ten minutes. Additionally, a CCD camera records fluorescent signals created by hybridization as the experiment is occurring, enabling the researchers to look at the total pattern of hybridization over a period in which temperature can be varied, including those that occurred earlier or at a different temperature.
The porous array material also enables PamGene to operate without infringing on Affymetrix patents for oligonucleotide arrays, which specifically refer to a “smooth and impermeable substrate.”
PamGene is not the only company to offer the porous chip. Gene Logic’s new majority-owned spin-off Metrigenix, is also developing a “flow-through” chip made out of porous silicon and glass microchannels. Drew O’Bierne, CEO of the Gaithersburg, Md, company, says PamGene’s technology “is limited to using a particular type of membrane, which is a subset of our technology.” Additionally, he said, Metrigenix is doing gene expression profiling, not just gene signatures and microbial work.
But Kievits says his chip offers the advantage of uniform sample flow, while the Metrigenix chip enables fluid to flow unevenly through the channels, inhibiting uniform hybridization and lengthening hybridization time to an hour. “That’s a technical problem that comes from a physical aspect of our material vs. theirs,” said Kievits. “Ours is macrofabricated while theirs is microfabricated. Microfabrication is much more costly.”
PamGene is marketing its chips to pharma and biotech companies that want to run samples through large numbers of chips with hundreds, not thousands, of distinct probes. The technology is designed to fit into the microtiter plate systems and microplate stackers.
Currently, PamGene can manufacture 16 chips an hour, and plans to push throughput up to 96 per hour once the technology is accepted in biotechnology and pharmaceutical companies.
Destined for Diagnostics?
Eventually, PamGene aims to go for the holy grail of molecular diagnostics. Given the speed of hybridization, the low cost of materials, and what the company claims is a robust assay with a high signal-to-noise ratio, the technology is well-suited to become a diagnostic chip, Kievits said. “We are the only system in the microarray world that will make it to diagnostics.”
Other systems that involve in-situ synthesis of oligonucleotides, he said, are not robust enough since there is too high an error rate for oligo identification, and the US Food and Drug administration requires over a 90 percent purity of materials.
But PamGene does not plan to go it alone in the diagnostic arena. With a decade of diagnostics experience under his belt, Kievits says he knows a small company is ill equipped to jump through regulatory hurdles, as well as setting up large-scale multi-product distribution channels in hospitals and clinical labs.
Luckily, PamGene may have a potential partner in parent company Organon Teknika, which Akzo Nobel sold last July to French diagnostics company Biomerieux. “That’s the new party to talk to,” Kievits said.