Method for reducing bacterial endotoxin contamination in solutions of macromolecules
AssigneeScripps Clinic and Research Foundation
InventorsThomas E. Karplus, Richard J. Ulevitch, Curtis B. Wilson
Life of this patent
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Abstract
The present invention contemplates a method of reducing a bacterial endotoxin contaminant in a biologically useful macromolecule. AN aqueous medium containing an endotoxin-contaminated macromolecule is admixed with a dialyzable surfactant, and the admixture so formed is contacted with an endotoxin sorbant to form a solid-liquid phase admixture. The contacting is maintained until the endotoxin is bound to the sorbant. The surfactant is dialyzed out of the aqueous liquid phase at a time no earlier than the maintenance step. The liquid phase containing the macromolecule is separated and recovered.
Claims27
A method of reducing a bacterial endotoxin contaminant in an aqueous composition containing a biologically useful macromolecule comprising the steps of:
- (a) admixing an endotoxin-contaminated biologically useful macromolecule with a dialyzable surfactant in an aqueous medium to form an aqueous admixture, said surfactant exhibiting no net electric charge at the pH value of said aqueous admixture and being present in said admixture at a concentration greater than said surfactant's critical micelle concentration;
- (b) contacting said aqueous admixture with a water-insoluble solid phase endotoxin sorbant comprising a solid phase matrix linked to an endotoxin sorbing agent to form a solid-liquid phase admixture;
- (c) maintaining said contact for a time period sufficient for said endotoxin to bind to said sorbant and form a second solid-liquid phase admixture whose liquid phase contains water, said macromolecule and a reduced weight ratio of endotoxin to macromolecule as compared to the weight ratio present in said aqueous admixture;
- (d) separating the solid and liquid phases of said second solid-liquid phase admixture;
- (e) dialyzing said surfactant from said liquid phase at a time no earlier than step (c) to provide a liquid phase that contains said macromolecule and is substantially free of surfactant;
- and
- (f) thereafter recovering the liquid phase that is substantially free from surfactant.
A method of reducing a bacterial endotoxin contaminant in an aqueous composition containing a biologically useful macromolecule comprising the steps of:
- (a) admixing an endotoxin-contaminated biologically useful macromolecule with a surfactant in an aqueous medium to form an aqueous admixture, said surfactant exhibiting no net electric charge at the pH value of said admixture and being present in said admixture at a concentration greater than said surfactant's critical micelle concentration;
- (b) contacting said aqueous admixture with a water-insoluble solid phase endotoxin sorbant comprising a solid phase matrix linked to an endotoxin sorbing agent to form a solid-liquid phase admixture;
- (c) maintaining said contact for a time period sufficient for said endotoxin to bind to said sorbant and form a second solid-liquid phase admixture whose liquid phase contains water, said surfactant, said macromolecule and a reduced weight ratio of endotoxin to macromolecule as compared to the weight ratio present in said aqueous admixture;
- (d) separating the solid and liquid phases of said second solid-liquid admixture;
- (e) dialyzing the surfactant out of said separated liquid phase to provide a liquid phase that is substantially free of surfactant;
- and
- (f) recovering said substantially surfactant-free liquid phase.
A method of reducing a bacterial endotoxin contaminant in an aqueous composition containing a biologically useful macromolecule comprising the steps of:
- (a) admixing an endotoxin-contaminated biologically useful macromolecule with a surfactant in an aqueous medium to form an aqueous admixture, said surfactant exhibiting no net electric charge at the pH value of said admixture and being present in said admixture at a concentration greater than said surfactant's critical micelle concentration;
- (b) contacting said aqueous admixture with a water-insoluble solid phase endotoxin sorbant comprising a solid phase matrix linked to an endotoxin sorbing agent to form a solid-liquid phase admixture;
- (c) maintaining said contact while dialyzing said solid-liquid phase admixture for a time period sufficient for said endotoxin to bind to said sorbant and form a second solid-liquid phase admixture whose liquid phase contains water, said macromolecule, a reduced weight ratio of endotoxin to macromolecule as compared to the weight ratio present in said aqueous admixture, and is substantially free of said surfactant;
- (d) separating the solid and liquid phases of said second solid-liquid admixture;
- and
- (e) recovering said separated liquid phase.
Dependencies are inferred from the claim text.