Patent ID: 8084581

Claim:
A method for the preparation of an oxygen carrier-containing pharmaceutical composition, the oxygen carrier-containing pharmaceutical composition including hemoglobin, the hemoglobin including cross-linked polymeric hemoglobin, the method comprising: (a) providing whole blood including at least red blood cells and plasma; (b) separating the red blood cells from the plasma in the whole blood; (c) filtering the red blood cells that were separated from the plasma to obtain a filtered red blood cell fraction; (d) lysing the red blood cells to create a solution comprising a lysate of disrupted red blood cells; (e) extracting a first hemoglobin solution from the lysate; (f) performing one or more purification processes on the first hemoglobin solution; (g) cross-linking hemoglobin tetramers in the first hemoglobin solution to create a second hemoglobin solution including cross-linked polymeric hemoglobin, the polymeric hemoglobin including two or more cross-linked hemoglobin tetramers; (h) heat treating the second hemoglobin solution including the cross-linked polymeric hemoglobin in a deoxygenated environment at a temperature greater than 85° C. and less than or equal to approximately 95° C. for a period of less than approximately 40 minutes to denature and precipitate any residual non-reacted hemoglobin, non-stabilized hemoglobin (dimer) and any other impurities; (i) cooling the heat-treated solution to a temperature approximately less than or equal to 25° C. in approximately two minutes or less to prevent formation of met-hemoglobin; (j) removing precipitate by a centrifugation or a filtration apparatus to form a solution of hemoglobin including cross-linked polymeric hemoglobin.