Purification and chemical modifications of hemoglobin in developing hemoglobin based oxygen carriers

Purification and characterization of human hemoglobin: effect of the hemolysis conditions

“…Table 2 shows an estimation of phospholipids concentrations detected by HPLC for hemoglobin solutions, obtained from hemolysis in water and additional purification by anion exchange chromatography. Although cold water may be considered as the best hypotonic medium for hemolysis [26], dilution of hemoglobin solutions in pure water seems not to contribute to purification by anion exchange chromatography, at least on AG MP-1 and Q-SFF resins, since substantial amounts of PC were not retained by these resins. Both resin type and flow rate influenced the efficiency of the process.…”

High O2 affinity hemoglobin‐based oxygen carriers synthesized via polymerization of hemoglobin with ring‐opened 2‐chloroethyl‐β‐D‐fructopyranoside and 1‐o‐octyl‐β‐D‐glucopyranoside

“…Intact bRBCs were then lysed with three equivalents of hypotonic 15 mOsM phosphate buffer (Haney et al, 2000). Cell debris was removed by passing the lysed bRBC solution through a hollow fiber cartridge with a pore size of 0.05 mm.…”

Purification of bovine hemoglobin via fast performance liquid chromatography

“…Initial studies focused on the use of centrifugation to first wash the RBCs and remove plasma proteins followed by the addition of a hypotonic buffer to extract Hb from the RBCs [15]. This was sometimes followed by the addition of toluene or another organic solvent to remove cell debris; however, it was found that toluene could remain within a hydrophobic pocket in Hb [16].…”

“…Hb purification has also been performed with Q Sepharose Big Beads by Lu et al [21]. However, this method was performed at a pH 6.8 [21], which is of concern since metHb forms rapidly at a pH less than 7 [15]. Additionally, the Hb was not absorbed by the resin, which increases the amount of lipid in the final bHb product.…”