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The Warsaw Voice » Other » September 30, 2009
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Blood as a Cure
September 30, 2009   
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Every year, 2 million patients in Poland are treated with blood and blood products. But only blood compatible with the recipient's blood type can be safely transfused. The Jagiellonian University (UJ) in Cracow and the Regional Blood Donation and Treatment Center (RCKiK) in Katowice have set up a consortium to develop modern reagents for serological tests.

To finance the project, the consortium has applied for zl.1.5 million in funds from the Ministry of Science and Higher Education's IniTech program.

Dominik Czaplicki, a PhD at the Jagiellonian University's Innovation, Technology Transfer and Development Center, says that reagents now used for blood grouping are based on monoclonal antibodies that detect only one type of antigens. However, the reagents are not produced in Poland and have to be imported.

A research team headed by Assoc. Prof. Joanna Bereta, of the Jagiellonian University's Biochemistry, Biophysics and Biotechnology Department, is working to generate new lines of cells that produce monoclonal antibodies. The university has a specialist laboratory where the researchers can carry out projects of this kind. The Regional Blood Donation and Treatment Center in Katowice is Poland's only blood donation center producing serological reagents. The consortium of the two institutions aims to develop a technology for the production of blood grouping reagents. Although the technology will not be new by global standards, the project is expected to yield a set of reagents-cheaper than their imported counterparts-to identify every blood type (O, A, B and AB) and the D antigen, called the Rh factor.

Bereta is in charge of the research side of the undertaking. Dr. Katarzyna Rothkegel, head of the Serological Reagents Production Department at the Regional Blood Donation and Treatment Center in Katowice, and Dr. Stanisław Dyl±g, the center's director, are responsible for the commercial side of the project.

"The project is planned for four years-this much time is needed to develop the technology and launch production," says Czaplicki. "We hope that spending from the national budget will be recouped in five or seven years and that the research will produce savings for taxpayers."

The consortium wants to carry out the project in two stages. The first stage, research work, will be conducted in Cracow. The second stage will involve preparations to launch production. The reagents will be certified, standardized and approved. Then, the Regional Blood Donation and Treatment Center will begin to produce reagents in its laboratories in Katowice.

The project is expected to result in developing original intellectual property that will be protected as know-how. The new technology is designed to enable production of several new lines of cells producing antibodies. The final product will be a reagent containing antibodies rather than cells that produce them, Czaplicki says.

Preventing blood clots
Krzysztof Szczubiałka, PhD, and his team at the Nanotechnology of Polymers and Biomaterials Research Group at the Jagiellonian University's Faculty of Chemistry have been successful in work on new polymer materials for the removal and release of heparin. Heparin is a strong anticoagulant that is often used during surgical procedures. But if it is left in the patient's blood it may cause dangerous hemorrhages.

The researchers expect to receive zl.238,000 in EU structural funds for the protection of their inventions with international patents. The funds will come under the EU's Innovative Economy Operational Program, which, among other objectives, aims to help protect industrial property created by research and development centers.

Heparin is sold under various trade names, such as Fragmin, Fraxiparine and Lovenox/Clexane. It prevents the formation and expansion of blood clots. Heparin is most often used in cases of acute myocardial infarction and in cardiac surgery.

In all cases when it is necessary to prevent blood clotting, it is important to ensure a long-term and stable concentration of heparin in the blood. Heparin has a half-life of only one hour or so, which is its major drawback. The Cracow researchers want to prolong the time of heparin release.

Their first invention for which they are seeking international patent protection is a biomaterial that releases heparin in a controlled way for over 10 days. The biomaterial is based on a hydrogel called alginiate-hydroxypropyl cellulose. The release speed can be controlled by the composition of the material and by the temperature because the material is thermosensitive.

The material comes in the form of microcapsules, a film, or a film containing microcapsules. It may find application in the oral administration of heparin to patients and as scaffolding for blood vessel tissue engineering. The inventors are Anna Karewicz, Katarzyna Zasada, Krzysztof Szczubiałka, and Maria Nowakowska.

The Cracow researchers also worked to find a way for heparin removal from the blood circulation system. The clinical use of heparin is not limited to cardiac surgery. The drug is also used in other conditions when a fast anticoagulation effect is needed-for example during other surgical procedures and in medical equipment such as renal dialysis machines and oxygenators, in order to prevent blood clotting.

However, since heparin has many undesirable side effects, it is necessary to remove it from the circulation system after the required anticoagulation effect has been achieved. The method used most frequently is the administration of protamine sulfate, a drug that reverses the anticoagulant effect of heparin by binding to it. Unfortunately, in around 10 percent of patients, the drug causes undesirable reactions that may even lead to death.

Patent pending
Another invention that is waiting for patent protection abroad is a method for removing and neutralizing heparin in blood and saline solutions by means of a modified chitosan. The polymer can be used both in a soluble form and in the form of microspheres or a film. Chitosan is a natural, biodegradable and biocompatible polysaccharide. Its main advantages are low cost and non-toxicity. Genipin, which the researchers used for cross-linking, is a natural non-toxic compound found in the gardenia fruit extract.

The polymer can be used in two ways-either as a solution that can be quickly administered as an intravenous injection, for instance to reverse the anticoagulation effect of heparin, or in the form of microspheres or a film, which may find application in the production of devices for the extracorporeal removal of heparin from blood and plasma.

The chitosan production method developed by the researchers yields a product with the desired effect on heparin binding. Its additional advantage is its effective antibacterial and antifungal activity. The polymer can be easily reclaimed and used again to remove heparin.

The biomaterials developed by the researchers and the methods for removing and neutralizing heparin with their use are the subject of domestic patent applications. Additionally, Kamil Kamiński, Karolina Zazakowny, Krzysztof Szczubiałka and Maria Nowakowska, who headed the research project, have won plaudits from the American Chemical Society for their publication entitled pH-Sensitive Genipin-Cross-Linked Chitosan Microspheres for Heparin Removal.

"The strategy to internationally expand the patent applications in the proposed project involves the use of the international Patent Cooperation Treaty (PCT) procedure and applications to the European Patent Office and the United States Patent and Trademark Office," says Czaplicki.
Piotr Bartosz
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