"If you stare at a knothole long enough, you’ll see something that nobody else has ever seen."

Although he first heard that pearl of wisdom some 20 years ago, a Wayne State professor not only remembers it, but lives by it. Not long after he joined Wayne State’s pathology department, Joseph Artiss, PhD, heard that advice from fellow faculty member Dr. Bennie Zak, now professor emeritus.

Many things have changed since then, but for Dr. Artiss, who still works with Dr. Zak developing new assays (nine of which are already licensed), the words still ring true. "With so many of these things, you look at them and you see the same things everybody else has seen for the last 10, 12, 15 or 20 years," Dr. Artiss remarked. "Finally you say, ‘There’s got to be a better way.’"

Through a nearly 20-year, stop-and-go career as an assay developer, Dr. Artiss has worked with other researchers, both academic and industrial, and most recently with a small Michigan company to improve upon – sometimes dramatically – the standard diagnostic reagents available for clinical testing.

"A lot of people feel the (development of) basic chemical tests is a mature business," Dr. Artiss said. "For the most part, the big manufacturers have disassembled their development groups in this area. I think they feel as if they’ve invested as much as they want to invest, and they want to reap the benefits. It’s really too bad." This mentality left the door wide open for Artiss and the Lincoln Park company, Pointe Scientific Inc. Currently, he added, "We have three technology transfer agreements involving the nine reagents mentioned above, and another in preparation. These are all improvements on existing assays or reagents."

To substantiate his point, Dr. Artiss leafed through a trade magazine to find an advertisement for a piece of equipment that performs immunoassays. He explained that instead of developing assays that are more accurate and precise and probably less expensive, the company has simply repackaged old technology into a new box. "From a very short-sighted corporate perspective, I can see it," the outspoken professor said. "But the same interferences you were getting 10 years ago, you’re still getting today. The erroneous results you were getting 10 years ago, you’re still getting today. The only difference is you’re getting them faster, because you have a faster box. Is there an advantage to making more mistakes faster?"

At the request of Pointe Scientific, Dr. Artiss began investigating homogeneous immunoassays that would work on any clinical analyzer. Most current assays require separate, stand-alone units. After reading an article about a new type of binding assay, he approached Wayne Lancaster, PhD, and Russell Finley, PhD, of the WSU Center for Molecular Medicine and Genetics. After a number of conversations, Dr. Artiss said, "We came up with an idea for an approach on this homogeneous binding assay which is exactly what (Pointe) wanted. My approach is to work on the chemistry and make the chemistry work, then anybody can build a box to put it on." The concept for this assay is an encouraging one, he added. "It should work better, quicker, simpler, easier, and be less-costly. It is currently the subject of a small business technology transfer grant proposal."

The key to his success lies in his ability to look at the problems differently, or to "stare at a knothole long enough." Because he isn’t wedded to a particular technology, he said, "I don’t have to do things the way they’ve been done for 20 years. For a lot of the large companies, it’s easier and more economical to take the technology they’ve been using for 10 or 20 years and put it in a new box."

Instead of corporate-imposed restrictions, Dr. Artiss’s constraints are "accuracy, precision, reagent stability, lack of interferences – all the things they teach you are right." He also must consider the trickier aspects inherent to the assays, such as their stability under different environmental conditions. "I need to think about India in the hot season. Enzymes are proteins; they will denature. On the other hand, you’re going to have to ship that same reagent to Minnesota in January. What does the cold do to it?"

An example is the reagent used to detect kidney disease. "The most common reagent for the determination of creatinine was developed in the late 1800s, and we’re still using that technology today," he said. The problem is that the reagent contains detergent, which precipitates when stored at cold temperatures. Although the precipitate goes back into solution if it is returned to room temperature, technologists often don’t have the luxury of waiting the few hours for the reagent to warm sufficiently.

Dr. Artiss took a new approach that eliminates the precipitate. "We worked out a way around it. It works very well."

He and Dr. Zak already have licensed calcium, iron-binding, creatinine, high-sensitivity cholesterol, triglyceride and four other assays. Once the project is completed and the problem solved, however, Artiss quickly shifts his interests elsewhere. He is particularly excited about current projects involving a new color indicator for liver disease and a quick, sensitive assay for viruses.

The color test, which he calls "ALT on a stick," changes colors at set concentrations of alanine aminotransferase, or ALT. An elevated level of this enzyme in the blood serum is a tell-tale sign of liver damage and perhaps hepatitis. "Our thought was to put it into a "dipstick" format to do spot tests of blood donors. If ALT is elevated, we can rule out that donor immediately," he said. "It will save the blood banks – the Red Cross – a lot of money, because they won’t have to do any further testing, or collect the donation, screen it and then destroy it. They can just say, ‘Thank you very much. You’d best go see a physician.’"

The development of ALT on a stick unfolded when he and friend Keith Taylor, a chemist and biochemist at the University of Windsor, met with a company’s representatives to do some contract work. The company wanted an indicator that would change colors, not just intensities of a single color. "He and I looked at each other ... and thought, ‘This is impossible.’ But we took their money, and a couple of months later had something that did just that."

This technology, which Taylor and Artiss created, involved straightforward chemistry, but was exhilarating nonetheless, Dr. Artiss said. To create the indicator, they began with two reduced substrates, one colored and one colorless. Dr. Taylor then synthesized a series of substrates that would preferentially oxidize from a colored to a colorless compound. Once the colored substrate was gone, the same enzyme would begin oxidizing the colorless substrate, which made it magenta. In the end, they were able to adjust the indicator so that at set substrate concentrations, it would switch from peach to clear to pink and finally to magenta. This novel color reaction forms the basis for the new test.

"For somebody who knows what’s going on in the test tube, you can’t help but smile when you see it happening, because it’s textbook classic enzyme kinetics, but you just wouldn’t expect it to happen," Artiss said.

Another project that holds his attention at the moment is barely off the drawing board. He is part of a team of researchers, including an engineer, working on a virus-detection assay. "What we think we can do is measure as few as 10-100 virus particles in a matter of a few minutes, which would be a very big step forward in technology for clinical laboratories. Not only could you measure this, but with a simple washing system – just passing buffer through – the system would be ready for a second sample."

Although technology known as PCR, or polymerase chain reaction, is even more sensitive, it involves amplifying the virus particles, which is time-consuming. "We’re thinking we can take procedures that are now taking 24-48 hours and reduce them to 15-30 minutes. (With our system), you would be able to significantly reduce materials and labor costs," Dr. Artiss said, reiterating that the project is still in its preliminary stages.

The idea for the virus-detection assay dates back to fall 1997 when Dr. Wayne Lancaster of the Center for Molecular Medicine and Genetics (CMMG), and George Grunberger, MD, of internal medicine and director of the CMMG, introduced Artiss to the work of Paul Van Tassel, PhD, a WSU professor of chemical engineering and material science, and specialist in surface chemistry. Artiss met with Van Tassel, who explained that his system could detect and quantify the presence of molecules deposited on a solid surface.

Artiss left the meeting sure the system had an application in clinical testing, probably immunoassays, but couldn’t pinpoint it until the following day when he was back on the medical campus. "I was walking across the yard out here, and it dawned on me: Everybody and his brother has an immunoassay system; what we want is to detect nucleic acids. Go after virus particles."

From there, a team developed. Dr. Lancaster does the nucleic acid work; Dr. Taylor from the University of Windsor does the immobilization work, which involves attaching the nucleic acids to a solid surface; Dr. Van Tassel does the optics work, which detects and counts the virus particles; and Dr. Artiss lends his expertise to ensure that the final product is a worthy addition to a clinical setting.

As of the summer of 1998, the team was busy with optimizing the immobilization process. Dr. Artiss said they are also working with both a synthetic polymer of nucleic acid and a real virus nucleic acid strand. "Our approach is to have both a synthetic and a real sample as proof of the principle. With that, we can do correlation studies with existing technology to prove that what we’re doing is what we think we’re doing."

Once these projects are completed, Dr. Artiss will no doubt turn his attention to yet another assay that he thinks needs improvement. "It has to be a problem. I like solving problems," he explained. "I think that a lot of people sort of become complacent with the status quo. I like to look for the opportunities in the status quo."

Another day, another knothole.