Method for preparation of thrombin
AssigneesOmrix Biopharmaceuticals Ltd., Guangzhou Bioseal Biotech Co., Ltd., Guangzhou Science Co., Inc.
InventorsLior Weissman, Dana Sella, Israel Nur, Caixia Jiang, Baochang An
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Abstract
The present invention is directed to methods for the production of thrombin from a source of prothrombin using a given BaSO4 reagent as an adsorbent of prothrombin as well as methods for evaluating the suitability of a given BaSO4 reagent for use in preparation of thrombin. In at least one embodiment, the method includes contacting a sample of the given BaSO4 reagent with a source of prothrombin to obtain BaSO4-adsorbed prothrombin, and subsequently evaluating the pro-coagulant activity of the BaSO4-adsorbed prothrombin. In another embodiment, the method includes the step of evaluating relative to the pro-coagulant activity of normal mammalian plasma.
Claims18
A method for the preparation of thrombin from a source of prothrombin, the method comprising:
- a. providing one or more BaSO4 reagents and a source of prothrombin;
- b. contacting the one or more BaSO4 reagents and the source of prothrombin under conditions allowing adsorption of prothrombin with the one or more BaSO4 reagents, thereby obtaining BaSO4-adsorbed prothrombin;
- c. testing the pro-coagulant activity of the BaSO4-adsorbed prothrombin, and testing same pro-coagulant activity of normal mammalian plasma source;
- d. evaluating the one or more BaSO4 reagents as suitable BaSO4 adsorbing reagent and selecting a suitable reagent based on said evaluation;
- wherein the evaluation is carried out by comparing the pro-coagulant activity of the BaSO4-adsorbed prothrombin to the same pro-coagulant activity of a normal mammalian plasma source, the selection of suitable BaSO4 reagent for use in preparing thrombin is based on the pro-coagulant activity of the BaSO4-adsorbed prothrombin being not greater than the pro-coagulant activity of normal mammalian plasma source;
- e. contacting the suitable BaSO4 with a source of prothrombin under conditions allowing prothrombin adsorption;
- f. eluting a prothrombin-containing fraction from the BaSO4-adsorbed prothrombin by contacting the adsorbed prothrombin obtained in step e. with an elution buffer;
- and
- g. subjecting the eluted fraction to conditions which allow conversion of prothrombin into thrombin, thereby obtaining thrombin.
A method for evaluating the suitability of one or more BaSO4 reagents for use as a prothrombin adsorbent in preparing thrombin from a source of prothrombin, the method comprising:
- a. providing one or more BaSO4 reagents and a source of prothrombin;
- b. contacting the one or more BaSO4 reagents and the source of prothrombin under conditions allowing adsorption of prothrombin to the one or more BaSO4 reagents, thereby obtaining BaSO4-adsorbed prothrombin;
- c. testing the pro-coagulant activity of the BaSO4-adsorbed prothrombin and testing same pro-coagulant activity of normal mammalian plasma source;
- and;
- d evaluating the one or more BaSO4 reagents as suitable BaSO4 adsorbing reagents and selecting those suitable reagents based on said evaluation;
- wherein the evaluation is carried out by comparing the pro-coagulant activity of the BaSO4-adsorbed prothrombin to the same pro-coagulant activity of a normal mammalian plasma source, the selection of suitable one or more BaSO4 reagents for use as a prothrombin adsorbant in method of preparing thrombin is based on the pro-coagulant activity of the BaSO4-adsorbed prothrombin being not greater than the pro-coagulant activity of normal mammalian plasma source.
Dependencies are inferred from the claim text.