By Debbie Strickland

Special To BioWorld Today

Third Wave Technologies Inc. raised $19.5 million to fund scale-up and further development of a nucleic acid analysis methodology that the company is positioning to compete with widely used polymerase chain reaction (PCR) technology.

"Almost all the approaches today use PCR and rely on PCR primers to hybridize DNA and RNA to generate specificity," said Lance Fors, president and CEO of the privately held Madison, Wis., company. "Our technology simplifies the process by not relying on hybridization, but on a simple enzyme-substrate reaction. It's simpler and more specific, and allows us to develop high-quality assays faster and less expensively."

Dubbed Invader, the technology uses two nucleic acid probes, one of which is recognized and cleaved by an enzyme, to identify a specific sequence of DNA or RNA. The presence of the released cleaved probe then can be measured using existing read-out devices such as microtiter plate readers, mass spectrometry and DNA chips.

The method springs from an "original observation" made at the University of Wisconsin but, said Fors, "99-plus percent of the technology was developed [at Third Wave]."

Applications of the technology include genomic target discovery and direct clinical use of assays in infection and genetic testing. The company already has ventured into genetic predisposition testing in the cardiovascular field, targeting the nation's top 200 molecular diagnostics labs with its test for the Factor V (Leiden) mutation, which is linked to deep-vein thrombosis.

"Our goal is to introduce 10 clinical diagnostics products over the next 12 months," Fors said. While the costs of tests will likely vary, the Factor V test averages $22 per patient, he said.

On the research side, Invader has attracted the interest of various sponsors and research centers involved in the SNP Consortium, a non-profit organization launched in April to map single nucleotide polymorphisms in humans. SNPs are common variations in DNA. Locating them is expected to help researchers find genetic differences that predispose some people to certain diseases and that underlie much of the variability of response to drugs. (See BioWorld Today, April 16, 1999, p. 1.)

The Wellcome Trust, a $14 million consortium sponsor, was among the investors in Third Wave's latest financing, and the company has collaborated with academic sites affiliated with the consortium, including the Sanger Centre in Cambridge, U.K., and the Stanford Genome Center in Palo Alto, Calif. John Todd, a researcher at the Cambridge Institute of Medical Research, said in a prepared statement that Third Wave's platform "could potentially become the standard for the detection of genetic mutations and SNPs. The Invader technology represents the next generation of technology after PCR."

Fors said Third Wave has worked with half of the consortium's 10 big pharma sponsors.

Next Financing Could Be An IPO

The latest round of financing will be used to fund automation of assay development and manufacturing, allowing the company to scale up to meet anticipated demand. New investors in the financing include Schroder Ventures International, Life Sciences Fund II, The Wellcome Trust, SR One and MVM Ltd. Previous investors who returned include Venture Investors of Wisconsin and Kenneth McGuire.

Third Wave has raised $38 million over the course of its five-year history and is nearing an initial public offering, Fors said.

"We've sort of been below the radar screen on purpose," he said. "But the technology is here today - it's been validated by most of the major pharmaceutical companies and genome centers. Now we're targeting an IPO [initial public offering] in the next one to two years to help us get to the next level of critical mass."

Third Wave's business plan is multi-pronged. In addition to making and marketing diagnostic products, the company plans to develop assays and conduct target research both in house and in collaboration with partners, and to out-license its technology widely on a non-exclusive basis.

"This could essentially become an operating system," Fors said. "We want to enable people to use it."

In addition to Invader, the company has developed a platform called Cleavase Fragment Length Polymorphism (CFLP) Analysis designed to compete with sequencing analysis of nucleic acids. The technology uses the Cleavase enzyme to split junctions between single-stranded and double-stranded DNA to create collections of fragments for analysis.