Convenient method to purify hemoglobin.
A convenient method to purify Hb solution from outdated RBC has been established for the starting material of Hb-based blood substitutes. To prevent MetHb formation during the procedure, Hb in RBC was carbonylated in advance. Then RBC was mixed with organic solvent for hemolysis and centrifuged for removal of stroma. The resulting SFHb solution was heated at 60 degrees C and generated precipitates were removed out by centrifugation. The purity of Hb (25 g/dl) was confirmed by SDS-PAGE. IEF and oxygen binding property of the Hb solution also guaranteed its purity and no denaturation of Hb. This method is applicable to large scale production of the purified Hb for the starting material of Hb-based blood substitutes.
Sakai
H
H
Department of Polymer Chemistry, Waseda University, Tokyo, Japan.
Takeoka
S
S
Nishide
H
H
Tsuchida
E
E
D001802:Blood Substitutes / Q000737:chemistry / Q000302:isolation & purification*; D002263:Carboxyhemoglobin / Q000302:isolation & purification; D004912:Erythrocytes / Q000737:chemistry; D006454:Hemoglobins / Q000737:chemistry / Q000302:isolation & purification*; D006461:Hemolysis; D006358:Hot Temperature; D006801:Humans; D008722:Methods; D010108:Oxyhemoglobins / Q000302:isolation & purification; D011489:Protein Denaturation; D012997:Solvents
D001802:Blood Substitutes; D006454:Hemoglobins; D010108:Oxyhemoglobins; D012997:Solvents; C016484:hemoglobin, stroma free; D002263:Carboxyhemoglobin