Method for separating unmodified hemoglobin from cross-linked hemoglobin
AssigneeBiopure Corporation
InventorsWilliam R. Light, Maria S. Gawryl, Anthony J. Laccetti, Robert A. Houtchens
Life of this patent
Estimating the term…
Abstract
A method for separating unmodified hemoglobin from cross-linked hemoglobin in a hemoglobin solution. The method involves contacting the hemoglobin solution with a least one dissociating agent to form a dissociation solution wherein unmodified tetrameric hemoglobin is dissociated to form hemoglobin dimers. The hemoglobin dimers are then separated from the dissociation solution, while retaining the cross-linked hemoglobin in the dissociation solution.
Claims20
A method for separating unmodified hemoglobin from cross-linked hemoglobin in a hemoglobin solution, comprising the steps of:
- a) contacting the hemoglobin solution with a least one dissociating agent to form a dissociation solution wherein unmodified tetrameric hemoglobin is dissociated to form hemoglobin dimers;
- and
- b) separating the hemoglobin dimers from the dissociation solution, while retaining the cross-linked hemoglobin in said dissociation solution.
A method for separating unmodified hemoglobin from cross-linked in a hemoglobin solution to produce a physiologically acceptable blood-substitute, comprising the steps of:
- a) contacting the hemoglobin solution with a dissociating agent to form a dissociation solution, wherein unmodified tetrameric hemoglobin dissociated to form hemoglobin dimers;
- b) filtering the hemoglobin dimers from the dissociation solution while retaining cross-linked hemoglobin in the retentate;
- and
- c) washing the cross-linked hemoglobin with a physiologic buffer to produce a physiologically acceptable blood-substitute.
A method of claim 19 wherein the buffer contains sodium lactate and N-acetyl-L-cysteine.
A method for separating unmodified hemoglobin from cross-linked hemoglobin in a hemoglobin solution, comprising the steps of:
- a) mixing the hemoglobin solution with a dissociation agent, a buffer and a stabilizing agent to form a dissociation solution, wherein mixing is with low shear, and wherein the unmodified hemoglobin dissociates to form hemoglobin dimers within the dissociation solution;
- b) filtering the dissociation solution to separate the hemoglobin dimers from the cross-linked hemoglobin;
- and
- e) washing the cross-linked hemoglobin solution with a deoxygenated solution containing sodium lactate, N-acetyl-L-cysteine and physiologic electrolytes.
Dependencies are inferred from the claim text.