title: Use of the labscale tangential flow diafiltrator to remove tetrameric hemoglobin from polyhemoglobin, purify hemolysate, and concentrate polyhemoglobin.
abstract: The Labscale tangential flow diafiltration system in conjunction with polyethersulfone Biomax filters (both purchased from Millipore Inc.) were used in the following studies: (1) A crucial step in the preparation of polyhemoglobin (polyHb) is the removal of tetrameric hemoglobin (Hb), which can cause adverse side-effects. The efficiency for this depends on the integrity of the 100kDa filters. Those with lower integrity were not effective whereas those with a higher integrity of 0.075ml/min were more effective. A filter with an integrity of 0.075ml/min can reduce the initial 11.2% tetrameric Hb concentration to 1.3% using a flow rate of 0.2ml/min. Higher flow rate of 2.2ml/min was not as effective. (2) Filtration with a 100kDa filter was quick and efficient in separating stroma-free hemoglobin solutions from cellular debris from hemolysate. (3) Both 5kDa and 100kDa filters are efficient in concentrating hemoglobin solutions.
authors.affiliation: Artificial Cells & Organs Research Centre, Department of Physiology, McGill University, Montreal, Quebec, Canada.
authors.forename: Ada
authors.initials: A
authors.lastname: Stefanescu
authors.forename: Thomas Ming Swi
authors.initials: TM
authors.lastname: Chang
mesh_terms: D001802:Blood Substitutes / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D005374:Filtration / Q000295:instrumentation / Q000379:methods*; D006454:Hemoglobins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D011108:Polymers / Q000737:chemistry / Q000378:metabolism*; D055503:Protein Multimerization; D012106:Research / Q000295:instrumentation; D012107:Research Design; D013450:Sulfones / Q000737:chemistry / Q000378:metabolism*
chemical_list: D001802:Blood Substitutes; D006454:Hemoglobins; D011108:Polymers; D013450:Sulfones; C037158:polyhemoglobin; C022840:polyether sulfone