Method of producing a fibrin monomer using a biotinylated enzyme and immobilized avidin

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Patent number: US6046024
Publication date: 2000-04-04
Inventor: BURTON STEVEN JAMES (GB); PEARSON JAMES C (GB); EDWARDSON PETER A D (GB)
Applicant: SQUIBB & SONS INC (US)
Classification:
- international: A61K5/00; C12N11/02; C07K17/00; G01N33/53
- european: A61L2/00P2F; A61L24/10F; C07K1/22; C07K14/36; C07K14/465; C07K14/75; C07K17/10; C12P21/06
Application number: US19970904369 19970801
Priority number(s): US19970904369 19970801; US19950556244 19951109
Abstract of US6046024

Compositions and methods for avidin immobilized on an inert support material, e.g. agarose, are disclosed. The compositions have high activity levels of avidin and may further include a bulking agent, e.g., maltose, and a protectant to maintain the stability and integrity of the avidin agarose during lyophilization and terminal sterilization processes. These compositions have applicability in any instance where avidin agarose and/or the avidin/biotin technology are useful. In particular, the present compositions are useful in an enzyme capture system to prepare fibrin monomer useful for fibrin sealants. The fibrin is prepared by subjecting a fibrinogen-containing composition to a biotinylated enzyme to convert the fibrinogen in the composition to a corresponding fibrin monomer. A fibrin monomer/biotinylated enzyme mixture is formed. A material comprising an avidin/inert support composition having 1000 biotin binding units of activity per gram of the composition, a bulking agent and a protectant, is then introduced into the enzyme mixture to form a complex of the material and the biotinylated enzyme. The complex and enzyme are then removed therefrom to recover a fibrin monomer. Bulking agents selected from nonionic water soluble compounds are used.
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