By Frances Bishopp

Staff Writer

It's a fishy story and A/F Protein Inc. has the rights to it.

The Waltham, Mass. company is developing the commercial potential of antifreeze proteins (AFP), which are found in the blood of certain cold-water fish and, according to Elliot Entis, co-founder, president and CEO of A/F Protein, highly effective in preserving biological materials at cold or freezing temperatures that would normally be destructive to those materials.

This ability allows for improved hypothermic and cryogenic preservation of a wide range of mammalian cells, tissues and whole organs.

AFPs also can be used, Entis said, under appropriate circumstances to enhance cell destruction at freezing temperatures, a characteristic with application in the cryosurgical treatment of tumors.

"We are the world's only commercial producer of antifreeze proteins," Entis said.

This development-stage biotechnology company, which also has a protein purification and research and development laboratory at St. John's, Newfoundland, Canada, has a total of nine patents issued or in application status, two of which are critical to the company. The first is for the use of antifreeze proteins for the preservation of mammalian cells, tissues and organs and the second is for the AguAdvantage gene technology, which is the transgenic technology used to create fast-growing fish.

In addition to protein applications, the regulatory sequences controlling expression of AFPs in fish have been used by the company to re-engineer the fish growth hormone gene, resulting in the development of AquAdvantage Bred transgenic salmon, which grow, Entis said, at a rate of 400 percent to 600 percent faster than other farm-raised salmon, offering significant economic advantage in commercial aquaculture.

A/F Protein has a separate business division, a fish hatchery, Aqua Bounty Farms on Prince Edward Island, Canada, to develop and market AquAdvantage fish.

In the last two years, the company has sold more than $600,000 worth of AFPs to firms and researchers worldwide for the purpose of product development. The company, Entis said, is working also to develop products that will be used to preserve animal embryos, human oocytes (unfertilized eggs), food products and to enhance the protective capabilities of skin-care products.

Another research area for the company is in cold preservation of platelets using the antifreeze proteins. The use of platelet transfusion for the prevention or treatment of bleeding associated with severe thrombocytopenia has increased considerably during the past decade.

Most platelet units transfused are administered prophylactically to reduce morbidity associated with hemorrhage in cancer patients undergoing myelosuppressive treatment. The need for platelets continues to increase, Entis said, based on greater use of intensive radio- and chemotherapeutic regimens that cause prolonged bone marrow aplasia.

Two challenges are faced in meeting this growing need: to improve treatment for a large number of cancer patients who become refractory to multiple transfusions and to develop more effective means of managing platelet units collected.

Platelets cannot be stored in vitro at temperatures below 22 degrees centigrade without significant loss of function due to cold-induced activation or for more than five days without increased risk of transfusion-associated infection, Entis explained.

Scientific studies, he continued, have shown that AFPs may provide a potentially cost-effective means of preserving platelet function in vitro at reduced temperatures for extended periods of time, at least 21 days, Entis said.

"Platelets can't be stored except at room temperature," Entis said. "If you store them below room temperature, effectively they lose their function when they are transfused back into the human body.

"This is an expensive problem up and down the supply line," Entis said, who added the company now is in the middle of in vivo testing of AFPs affects on platelets.

A/F Protein already has licensed the AquAdvantage technology to several firms in the U.S., Europe and New Zealand and has an alliance with a large corporate food supplier, which cannot be identified at this time, Entis said.

"We are not interested in supplying end-use products ourselves," Entis said. "Our strength is to develop uses for the antifreeze protein and then to work with corporate partners who have products that can make use of the antifreeze protein. We are looking for corporate partners in the area of cryosurgery and blood platelet preservation."

A/F Protein, founded in 1992, is preparing for a mezzanine financing, and has already raised $5.2 million of private capital through two rounds of Registration D financing, the second of which raised $4 million. *