08 Mar America’s Third Coast nurtures biotechnology
Wisconsin Offers Diverse Enterprises
The biotechnology community in Wisconsin calls itself “America’s Third Coast,” referring to its proximity to Lakes Superior and Michigan. Approximately 150 companies belong to the Wisconsin Biotechnology Association (WBA), based in Madison, which is also home to the Big 10 campus of the University of Wisconsin. With its worldwide reputation for scientific research, UW-Madison attracts an average of $600 million annually in outside research funds.
Hundreds of patents have been translated into start-up businesses, aided by WARF (Wisconsin Alumni Research Foundation). Since being founded in 1925, the private, nonprofit WARF patents inventions arising from university research, licenses the technology for commercialization, and returns the income to UW-Madison to support further research. WARF has contributed $650 million to UW-Madison over the years.
“The biotechnology industry is primarily in Madison, with some companies in Milwaukee,” says Jim Leonhart, executive vp of WBA. A main reason is that biotechnology companies in Madison are nurtured at the University Research Park, where they turn life science research into new drugs, therapies, and medical devices.
About 100 companies, employing 4,000 people, reside in the Park. Most have ties to UW-Madison and hold WARF patents. The Park’s 34 buildings cover 1.5 million sq ft, and a 13,000-sq-ft incubator site houses 35 smaller companies.
“After three years, we like to nudge them out of the incubator,” says Mark Bugher, director of the University Research Park. Core services, like accountants, lawyers, and venture capitalists are also on-site. A description of companies in the Park is listed at www.universityresearchpark.org.
Although the bioindustry is not as large or recognized as on the other two coasts, standouts like Promega (reagents and techniques for life science R&D; www.promega.com), Epicentre (molecular biology products for life science and clinical research, molecular diagnostics, and biopharmaceutical manufacturing; www.epicentre.com), Third Wave Technologies (molecular diagnostics; www.twt.com), and Scilog (bioprocess pumps and liquid handling systems; www.scilog.com) are known worldwide. Other biofirms representing the diversity of endeavors in Wisconsin are profiled below.
Keeping with Wisconsin’s nickname of the dairy state, Gala Biotech (www.gala.com) planned to produce pharmaceutical proteins in the milk of transgenic cows when it was founded six years ago. (“Gala” is Greek for milk.) But along the way, the company replaced cows with mammalian cell cultures. Gala’s core proprietary platform is GPEx™ (Gene Product Expression), a gene insertion/expression method that allows the rapid creation of stable, high-expressing mammalian cell lines for a variety of gene products.
GPEx relies on a safe version of the retroviral gene insertion machinery discovered by the late Howard Temin at UW-Madison, who won the Nobel Prize in 1975 for his discovery. “In biologics, there’s a huge shortfall in manufacturing methods for recombinant protein drugs,” says Paul Weiss, Ph.D., president of Gala Biotech.
Any recombinant protein can be manufactured by GPEx technology using mammalian cells growing in stainless steel tanks, the company says. Clients can contract with Gala Biotech to do the manufacturing at its 43,000-sq-ft protein production plant or license cell lines containing GPEx for manufacturing at their own site. In October 2003, Gala Biotech was acquired by Cardinal Health (Dublin, OH), which will assist in packaging and commercialization of products.
Technology developed in the laboratory of chemical engineer Nick Abbott, Ph.D., is being commercialized by Platypus Technologies (www.platypustech.com). Platypus uses liquid crystals, like those in computer monitors, to detect nanoscale changes on cell surfaces. The technology can detect viruses, bacteria, and environmental toxins like pesticides. Why the name Platypus? The platypus bill contains highly specialized receptors that detect prey in muddy water, and “our technology involves highly specialized surfaces to detect target molecules present in complicated mixtures,” explains virologist Barbara Israel, Ph.D., COO.
Researchers at Platypus attach liquid crystals to gold or polyurethane surfaces. When a virus or pesticide attaches, the alignment of the liquid crystals visibly changes and can be seen with the naked eye. With an SBIR grant from the National Institute of Environmental Health Sciences, Platypus is designing a detection system for organophosphonate pesticides.
The goal is to design a noninvasive, lightweight monitor to measure daily pesticide exposure without requiring blood tests. The same device could detect occupational exposure to similar compounds or nerve poisons like saran.
In addition, Platypus is developing prototype assays to detect viruses with liquid crystals. The detection system is rapid and sensitive, and no labels, probes, enzymes, or radioactivity are needed. “We can pattern a one-inch by two-inch surface to detect 20 different viruses,” says Dr. Israel. The probes can be used in diagnostic laboratories or in field work.
Mirus Technologies (www.genetransfer.com) offers nonviral gene transfection delivery products, which can be directly injected into the body to carry genes directly to target cells. Mirus was founded in 1995 by John Wolff, Ph.D., a UW-Madison pioneer in gene transfer, who holds some of the earliest patents on DNA delivery methods.
The company’s product line includes 10 transfection reagents for specific cell lines, three broad-spectrum products for plasmid delivery in multiple cell lines, specific reagents for transfection of oligonucleotides and siRNA, as well as labeling products for nucleic acids, plasmids, and fluorescent in situ hybridizations.
The next challenge is to develop intravascular delivery systems for delivering therapeutics in clinical trials. Mirus is collaborating with Transgene in France to test treatments for muscular dystrophy. Although Mirus is recognized as a niche provider of nucleic acid reagents, “our potential in therapeutic opportunities is under recognized,” says Russ Smestad, president of Mirus.
OpGen (www.opgen.com), uses a microfluidic technology, called the OptiChip™, to detect genetic variations in DNA samples without the need for DNA sequencing, amplification, and probe labeling. As a sample flows through multiple channels of the OptiChip, individual DNA molecules elongate and stick to the surface, forming a massive array that can be analyzed for specific markers. OptiChip captures not only genes, but also all of the DNA in between.
“We can detect gene arrangements, insertions, deletions, and variations not detectable by other methods,” says Colin Dykes, Ph.D., CSO. Although many researchers focus on SNPs, “there is growing evidence that insertions and deletions are more important,” he says. The company’s goal is to analyze human DNA samples for other companies engaged in clinical trials to find gene variations that correlate with drug responses.
OptiChip also analyzes the entire genomes of microbes that could be used as bioterrorist agents. Researchers at OpGen are constructing a database of infectious agents as a resource for rapid identification of suspect organisms. “Early identification of the causative agents in disease outbreaks is critical to mounting an effective response,” says Dr. Dykes.
Bone Care International (www.bonecare.com) specializes in novel vitamin D therapies for people with chronic kidney disease (CKD). The company’s main drug, Hectorol, treats secondary hyperparathyroidism (SHPT) in patients with CKD.
Healthy kidneys produce D hormone, which regulates calcium balance. Left untreated, low D hormone levels often trigger an imbalance causing parathyroid hormone levels to rise resulting in SHPT and, ultimately, the loss of calcium from bone, potentially leading to bone disorders and surgical removal of the parathyroid gland.
Vitamin D hormone levels are often impaired by chronic kidney disease. “We have one of three products on the U.S. market, Hectorol, to treat vitamin D deficiency in patients with chronic kidney disease,” says Basil Mundy, vp of corporate development.
Hectorol is given intravenously to dialysis patients. A capsule form of Hectorol is also available for peritoneal dialysis patients. Bone Care is waiting approval from the FDA for oral Hectorol to help patients with chronic kidney disease prior to dialysis. Mounting evidence suggests that vitamin D hormone may also help treat psoriasis, osteoporosis, and may slow tumor growth.
Madison’s biotech environment is so appealing, that EraGen Biosciences (www.eragen.com) moved its operations from Gainesville, FL, in 1999.
EraGen’s products are based on three proprietary platforms: AEGIS™ (An Expanded Genetic Information System) for molecular diagnostics, MasterCatalog™ for functional proteomics, and TACS™ (Target Assisted Combinatorial Synthesis) for drug discovery. These products can stand alone, but are also designed to work synergistically to speed both the creation of new diagnostics and the discovery process.
The AEGIS platform includes GeneCode, a real time, quantitative PCR-based method that uses iso-cytosine and iso-guanosine to simplify identification. MasterCatalog is a genome software analysis system that uses a patented genetic evolutionary technology for in silico protein prediction. The software predicts protein function by computing the evolutionary characteristics associated with 130 specific genomes.
GeneCode and MasterCatalog helped to develop an assay for SARS, which is used by public health agencies worldwide. “The SARS assay was created in a week, once the genome was made public last April,” says Dr. Hrusovsky. And TACS is a target-driven technology that simplifies the detection and identification of leads during drug development.
At Quintessence Biosciences (www.quintbio.com), researchers are preparing their Evade™ Technology for preclinical trials.
“Evade is a mammalian protein that is cytotoxic to cancer cells,” says Ralph Kauten, president of Quintessence Biosciences. The basis of the Evade technology involves ribonucleases that degrade RNA.
Because intact RNA is essential for life, ribonucleases can be toxic to cells, especially rapidly proliferating cancer cells. However, cells also contain natural ribonuclease inhibitors (RI), which bind ribonucleases to prevent cell destruction.
The Evade technology genetically modifies selected ribonucleases to prevent them from binding RI and losing potency inside targeted cancer cells. The selective toxicity of ribonucleases also makes them ideal for linkage to antibodies or growth factors for therapeutic delivery.
“We are exploring these kinds of partnerships with antibody companies,” says Kauten. Cancer drugs based on ribonucleases look economically feasible, since other companies have such drugs in Phase III trials, according to Kauten.
Although getting a later start, the Evade ribonucleases from Quintessence Biosciences may be better because they are derived from mammalian cells (humans or cows), whereas the ribonucleases in clinical trials elsewhere come from amphibians. In the long term, mammalian ribonucleases “may prove less risky,” says Kauten.
Novagen, a brand of EMD Biosciences (www.emdbiosciences.com), has expertise in protein expression, notes Kim Harris, senior director of North American sales operations.
Core customers range from life science researchers in academic settings to those in pharmas and biotechs developing new drugs. The company’s flagship pET system allows researchers to express a wide variety of proteins in bacterial cells in order to study interactions before moving on to human models used in drug discovery.
“The Novagen brand of EMD Biosciences has a solid reputation because of our expertise in molecular biology and our extensive tools for structural, functional, and display proteomics research,” adds Mark Lepinske, marketing communications, Novagen.
Novagen also sells specialty reagents to assist with protein expression. For example, the BugBuster family of extraction reagents disrupts cells and extracts relevant proteins gently and efficiently, according to Harris.
Whether purifying proteins from yeast, E. coli, or mammalian cells, liberating soluble proteins must be done carefully. Harsh mechanical methods, like sonication, can harm a protein’s structure and activity. The novel, detergent-based YeastBuster, BugBuster, and CytoBuster Reagents perforate cell walls without denaturing proteins, continues Harris. After a brief incubation, target cells are harvested by centrifugation and the protein is extracted.
Novagen also markets Robo-Pop™ purification kits. “RoboPop is a quantum ad-vance in the purification process, especially as it applies to high throughput, because it eliminates the centrifuge and permits the entire expression and purification process to be performed in a single well of a multiwell plate,” notes Lepinske.
Gilson (www.gilson.com) has been supplying standard laboratory tools, like Pipetman®, to laboratories worldwide for over 60 years. The firm’s knowledge of liquid-transfer methods is also incorporated various automated instruments and chromatography systems for drug discovery, proteomics, and genomics. In addition to manufacturing pipettes, Gilson produces HPLC systems, high-throughput robotic workstations, fraction collectors, solid-phase extraction instruments, and injectors.
Gilson recently introduced the new MALDILC™ System, a tool designed to perform nano, capillary, and micro HPLC with fraction collection directly sent to MALDI plates. The plated fractions can then be analyzed repeatedly by MALDI-TOF MS. The MALDILC System’s HPLC-coupled fraction collection simplifies complex tryptic digests, which provides more readable MS spectra. The MALDILC System automates what has been a historically manual procedure.
Lucigen (www.lucigen.com) supplies advanced molecular biology reagents and systems as well as related custom services. Products include cloning systems, competent cells, DNA End Repair, and enzymes. The company offers services such as custom cloning and NanoClone™ and GapFree™ library construction.
Takara Mirus Bio (www.takaramirusbio.com) is a newly formed joint venture between Japan-based Takara Bio and Mirus Technology. Takara Bio and Mirus have a relationship that goes back for some time, based on research collaborations and Takara’s distribution of Mirus’ research product line in Japan.
Takara Mirus Bio will distribute Takara Bio’s complete product line in North and South America, generating expanded U.S. visibility and availability of Takara Bio’s premium life science research products, notes Leslie Garland Miller, marketing and sales specialist at Takara Mirus Bio.
nPoint (www.npoint.com) designs, manufactures, and sells ultraprecision positioning and motion control devices that are capable of rapid, precise, and repeatable positioning and motion at the nanometer scale, according to a company official.
nPoint products were designed to be instrumental in resolving research and OEM instrumentation problems at the submolecular and nanoscale levels by providing motion in increments of as little as a fraction of a nanometer with high precision and speed.
nPoint’s founding technology was developed at the University of Wisconsin, Madison. The company’s standard products include systems capable of movement in single and multiple axes, the accompanying control systems and computer interfaces.
The firm’s nanopositioners are available in a variety of materials and in configurations capable of supplying ranges of motion across a full spectrum of requirements, explains a company spokesperson.
Based in Brown Deer, a Milwaukee suburb, Pel-Freez Clinical Systems (www.pel-freez.com) designs and manufactures serological and molecular biology products that support bone marrow and organ transplant procedures.
“Our products are used to find the proper match between donors and recipients,” says Frank Langley, COO. In June 2003, Oslo, Norway-based Dynal Biotech acquired Pel-Freeze. The combination will make us “one of the leaders worldwide designing and distributing serological test kits,” says Langley.
Langley also serves as president of the WBA. About two thirds of WBA member companies reside in Madison, 90 miles west of Milwaukee. Langley and others dream of building a research and development park in Milwaukee, similar to University Research Park in Madison.
The goal is to combine private sector companies with academic institutions on the same campus to nurture start-ups. “Milwaukee already is known as a good manufacturing town. We need to apply those skills to the biosciences,” says Langley.
Milwaukee is home to a number of universities, including Marquette, the Medical College of Wisconsin, and the Milwaukee School of Engineering. The Medical College of Wisconsin, an independent, private institution, “is especially strong in developing medical and imaging devices,” says William Hendee, Ph.D., vp of technology.
The Medical College of Wisconsin has a long-term relationship with General Electric Medical Systems. “We prototype a lot of their equipment,” says Dr. Hendee. No formal plans are yet in place for the research park, but the movement “is starting to get traction, and we’re getting business leaders involved,” says Langley.
One company already based in Milwaukee is Cambridge Major Laboratories (www.c-mlabs.com), which performs custom chemical synthesis services. The firm supplies biopharmaceutical and biotechnology companies with a line of reagents used in the synthesis of oligonucleotides, as well as speciality reagents, fine chemicals, and generic drugs.
With seven kilo-lab suites and a pilot plant, Cambridge Major Laboratories also can produce compounds in quantities ranging from grams up to a ton. “We are dynamic, flexible, and can tackle any kind of chemistry,” says Michael Major, Ph.D., president and CEO of the four-year-old business
Pierce Nucleic Acid Technologies (www.piercebc.com) also performs chemical manufacturing in Milwaukee. For more than 50 years, Pierce has specialized in the synthesis, modification, and labeling of nucleic acid-related products. The company’s expertise spans the production of amidites, cross-linkers, nucleotides, purification systems, and modifying enzymes.
“We’re the folks behind the scenes,” says Carolyn Stock, marketing communications manager. Bulk or customized products are produced in ISO 9001 certified manufacturing facilities to ensure product reproducibility and strict adherence to specifications.
“Our site produces materials in a variety of areas, including antisense technologies, interference RNA, and microarrays,” says Stock. Pierce Nucleic Acid Technologies is part of Perbio Science (Helsingborg, Sweden), which was recently acquired by Fisher Scientific International (Hampton, NH).
Carol Potera is a writer for Genetic Engineering News. This article originally appeared in the March 1, 2004 edition of Genetic Engineering News and was reprinted in WTN with the permission of Genetic Engineering News.