开环棉籽糖分子内交联血红蛋白及其戊二醛的二次交联

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-freehemoglobinSFHb)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65000的未聚合的O-Ra-Hb组分为22.6%,分子量为130000～600000的组分为76.9%,分子量为600000以上的组分仅为0.5%,达到了预期目的.

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 Firstly, Stroma-free hemoglobin(SFHb) was purified from outdated human blood.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 P50. 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%.