开环棉籽糖分子内交联血红蛋白及其戊二醛的二次交联

Stroma-free Hemoglobin Cross-linked with O-Raffinose and Glutaraldehyde

用高碘酸钠氧化制备开环棉籽糖 ( O-Raffinose) ,将其作为交联剂对脱氧态无基质血红蛋白 ( Stroma-free hemoglobin SFHb)进行修饰 .十二烷基磺酸钠 -聚丙烯酰胺不连续电泳 ( SDS-PAGE)结果显示 ,开环棉籽糖交联血红蛋白 ( O-Ra-Hb)与 SFHb相比抗解聚性有明显提高 ;琼脂糖 ( Sepharose)凝胶层析结果显示 ,SFHb和 O-Ra-Hb出峰时间和峰形大小基本相同 ,说明交联反应主要发生在分子内的亚基间 ,基本没有分子间交联产物 .用戊二醛 ( GDA)在优化条件下对 O-Ra-Hb进行分子间二次交联 ,SDS-PAGE结果显示 ,二次交联产物稳定性比 O-Ra-Hb有了进一步提高 ;Sepharose凝胶层析结果表明 ,产物中分子量为 65 0 0 0的未聚合的 O-Ra-Hb组分为 2 2 .6% ,分子量为 1 3 0 0 0 0～ 60 0 0 0 0的组分为 76.9% ,分子量为 60 0 0 0 0以上的组分仅为 0 .5 % 。

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 Firstly, Stroma-free hemoglobin(SFHb) was purified from outdated human blood.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%.