Chemistry: molecular biology and microbiology – Carrier-bound or immobilized enzyme or microbial cell;... – Enzyme or microbial cell is immobilized on or in an...
Patent
1988-11-14
1991-03-19
Naff, David M.
Chemistry: molecular biology and microbiology
Carrier-bound or immobilized enzyme or microbial cell;...
Enzyme or microbial cell is immobilized on or in an...
424 9461, 435180, 435181, 435206, 514 56, C12N 1114, C12N 1108, A61K 3754, A61K 31715
Patent
active
050010620
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to the field of heparinization of materials for the purpose of imparting thereto a heparin layer which inhibits in contact with blood the adhesion of thrombocytes and the adsorption of blood proteins. Thus, the article according to the invention is especially suited for applications where there is contact with blood, e.g. medical applications. It is true that the methodology of heparinizing a material for the above-mentioned purpose is previously known per se, but the present invention relates to a novel, alternative method of adhering the heparin to the substrate, via a special, novel type of a pre-adsorbed layer. In addition to the above-mentioned article the invention relates to a process for the preparation thereof as well as to the use of said article for medical applications.
BACKGROUND OF THE INVENTION
To accomplish blood compatibility for different materials in contact with blood one of the most important methods has been to heparinize the surface thereof. Thus, the heparin layer on the surface inhibits, as was mentioned above, the adhesion of thrombocytes and the adsorption of blood proteins. Furthermore, the heparin must be enzymatically active in the blood coagulation process, which calls for specific necessities as to molecular conformation and mobility relative to the surface.
Two main principles for the heparinization have previously been utilized. The first one is based on colloidal precipitation, e.g. through complex-formation between amphiphilic amines and heparin. The second one utilizes the possibility of covalently bonding the heparin to the surface. However, these known principles have some limitations which means that there is continuous research for alternative or improved methods for the heparinization.
SUMMARY OF THE INVENTION
The present invention relates to an alternative or improved technique for the heparinization of surfaces, which technique eliminates or at least reduces the limitations of the prior art while at the same time imparting thereto, at least for certain applications, additional advantageous properties which have not been obtainable by the previously utilized technique. More specifically we have found that a specific protein, viz. lysozyme, possesses unexpected affinity for heparin and gives an outstanding adhesion to different substrate surfaces. The unexpectedly good results which have been obtained by the protein according to the invention will be described more in detail below, but primarily it can be mentioned that a very good adhesion to metal surfaces has been obtained, which is material in which previously known methods have shown deficiencies.
Lysozyme is a protein that is present in low concentrations in blood. Already there is an interesting advantage since the invention is based on the utilization of a substance which is non-foreign to the human organism. In addition thereto another interesting property of lysozyme is its antimicrobial properties which impart to the novel heparinized surface an antimicrobial activity; a security factor in storage and handling.
More specifically the article according to the invention is characterized in that heparin or a heparin-based material is adhered or bonded to the substrate via a layer of lysozyme or a derivative thereof which is pre-adsorbed to said substrate.
As was indicated above the novel technique according to the invention has been shown to work especially well for metal surfaces, in which previously known heparinizing methods have shown limitations. However, the invention is also applicable to other substrates which are chosen per se in accordance with previously known techniques, i.e. primarily such substrates that have has previously been desired to heparinize for the purpose of imparting to the same improved properties in applications where there is contact with blood. Examples of such materials are polymeric materials and glass. As far as polymeric materials are concerned it should be noted that the present invention is especially interesting in conn
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Bergentz Sven E.
Larsson Kare V.
Lindblad Bengt L. T.
Camurus AB
Naff David M.
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