Stroma-free hemoglobin cross-linked with O-raffinose and glutaraldehyde

Red cell substitutes are group of oxygen carriers designed to temporarily replace transfused blood. In this paper, a kind of red cell substitutes based on polymerized hemoglobin were prepared and in vitro characteristics were studied. Firstly, Stroma-free hemoglobin (SFHb) was purified from outdated human blood. SFHb was intramolecularly crosslinked with O-Raffinose, prepared from the ring opening oxidation of raffinose. The intramolecular crosslinking between amino groups within 2, 3-DPG binding pocket contributes to high P-50. Then O-Raffinose-Hb was intermolecularly crosslinked with glutaraldehyde under the optimized crosslinking condition. The process of crosslinking was monitored by SDS-PAGE and Sepharose gel chromatography. The percentage of the component of polyhemoglobin with a molecular weight range of 130 000-600 000 was 76.9%.