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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

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【作者】 王雁峰潘继伦俞耀庭

【Author】 WANG Yan-Feng, PAN Ji-Lun *, YU Yao-Ting (Key Laboratory of Bioactive Materials of Ministry of Education, Nankai University, Tianjin 300071, China)

【机构】 南开大学分子生物学研究所生物活性材料教育部重点实验室南开大学分子生物学研究所生物活性材料教育部重点实验室 天津300071天津300071天津300071

【摘要】 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 % ,达到了预期目的

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 Firstly, Stroma-free hemoglobin(SFHb) was purified from outdated human blood.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

  • 【文献出处】 高等学校化学学报 ,Chemical Research In Chinese Universities , 编辑部邮箱 ,2004年10期
  • 【分类号】O621
  • 【被引频次】2
  • 【下载频次】171
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