title: Immobilized hemoglobin in the purification of hemoglobin-based oxygen carriers.
abstract: Chemically modified hemoglobins can be used as oxygen carriers in cell-free fluids provided that they have a low oxygen affinity and are stable towards dissociation into subunits. The latter species are undesirable because they are filtered rapidly through the kidneys, have renal toxicity and are characterized by a high oxygen affinity. A most important step in the preparation of hemoglobin-based oxygen carriers is therefore their purification from any dissociable material. Hemoglobin immobilized as alpha beta dimers on Sepharose lends itself naturally to this purpose as it is able to interact in a specific and reversible way with soluble alpha beta dimers. Hemoglobin affinity columns are very effective in the purification of cross-linked and pseudo-cross-linked human and bovine hemoglobin. The applicability of the technique is enhanced by the ease with which alpha beta dimers from different species cross-interact to yield hybrid alpha 2 beta 2 tetramers. It is shown that hemoglobin affinity columns may provide analytical information on the cross-linking reaction itself.
authors.affiliation: CNR Center of Molecular Biology, Department of Biochemical Sciences, University La Sapienza, Rome, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Chiancone
authors.forename: C
authors.initials: C
authors.lastname: Fronticelli
authors.forename: M
authors.initials: M
authors.lastname: Gattoni
authors.forename: B K
authors.initials: BK
authors.lastname: Urbaitis
authors.forename: E
authors.initials: E
authors.lastname: Bucci
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods*; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D006454:Hemoglobins / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D010100:Oxygen / Q000378:metabolism*; D051381:Rats; D013057:Spectrum Analysis
chemical_list: D003432:Cross-Linking Reagents; D006454:Hemoglobins; D010100:Oxygen