Study on preparation of poly-Carboxyhemoglobin by glutaraldehyde crosslinking / 中国输血杂志

ABSTRACT

【Objective】 To investigate the preparation conditions of hemoglobin-based oxygen carrier (HBOC) at room temperature without oxygen isolation and the quality of poly-COHb. 【Methods】 Using human cord blood hemoglobin as raw material and glutaraldehyde as crosslinking agent, the experimental group selected hemoglobin concentration, glutaraldehyde to COHb molar ratio, reaction time and reaction temperature to do four factors and three levels of orthogonal test (n=3), to determine the best matching conditions and prepare three batches of poly-COHb, continuously.In the control, 3 batches of poly-Hb were prepared under low temperature and oxygen isolation.The contents of superlarge molecules, degree of polymerization, average molecular weight, methemoglobin and P50 of crosslinked products were compared between the two groups. 【Results】 The optimal matching conditions of poly-COHb were as follows [Hb] 70g/L, the molar ratio of glutaraldehyde to COHb was 11∶1, the reaction time was 60 min, and the reaction temperature was 25℃.Comparison of poly-COHb and poly-Hb The superlarge molecular content (%) was 0.67±0.51 vs 0.60±0.01 (P>0.05); degree of polymerization (%) and average molecular weight (kD) were 84.25±0.99 vs 54.16±5.12, and 235.27±13.50 vs 97.62±6.57, respectively.(P<0.05); methemohemoglobin ratio (%) was 2.40±0.66 vs 2.47±0.46 (P>0.05); P50 (mmHg) was 6.37±0.30 vs 14.20±1.09 (P<0.05). 【Conclusion】 Poly-COHb can be prepared at room temperature without oxygen isolation.Compared with poly-Hb, poly-COHb prepared under the experimental conditions can improve the polymerization degree and average molecular weight, and greatly reduce the difficulty of preparation.