京尼平交联人胎儿血红蛋白的实验研究  

Experimental study of Genipin cross-linked hemoglobin isolated from fetal

作　　者：李燊[1] 张鸿辉 刘盼和 郝莎莎 王红[1] 刘嘉馨[1] 杨成民[1,3] 周文涛[1] LI Shen;ZHANG Honghui;LIU Panhe;HAO Shasha;WANG Hong;LIU Jiaxin;YANG Chengmin;ZHOU Wentao(Institute of Blood Transfusion,Chinese Academy of Medical Sciences&Peking Union Medical College,Chengdu 610052,China;Chengdu Jiaying Medical Products Co;Tianjin Union Biotechnology Development Co)

机构地区：[1]中国医学科学院北京协和医学院输血研究所,四川成都610052 [2]成都市佳颖医用制品有限公司 [3]天津协和生物科技发展有限公司

出　　处：《中国输血杂志》2023年第4期302-305,共4页Chinese Journal of Blood Transfusion

基　　金：中国医学科学院医学与健康科技创新工程项目“血液安全保障及新产品新技术研究”(2021-I2M-1-060);四川省科学技术厅项目“PolyCHb增敏宫颈鳞癌放射治疗的机制探索”(2021YJ0122)。

摘　　要：目的采用京尼平(Genipin,Gnp)对人胎儿血红蛋白(hemoglobin isolated from fetal,HbF)交联反应,预期目标为有效聚合率≥70%(分子量在64~600 KD)和平均分子量100~300 KD。方法以HbF为原料,Gnp为交联剂。选择血红蛋白浓度(HbC)、Gnp与HbF的摩尔比、反应时间和反应pH进行四因素三水平正交试验(n=3),确定较佳匹配条件。制备三批次京尼平交联人胎儿血红蛋白(Gnp-HbF)样品,检查有效聚合率、二聚体、平均分子量、高铁血红蛋白(MetHb)含量及P50。结果正交试验得出Gnp-HbF较佳匹配条件:HbC为80g/L、Gnp与HbF摩尔比为7∶1、反应时间18h和反应pH为8.5。本条件下Gnp-HbF样品有效聚合率为(73.03±2.03)%,二聚体为(26.97±2.03)%,平均分子量为(167.70±28.40)KD,高铁血红蛋白占比(2.60±0.17)%,P50为(13.13±1.02)mmHg。结论Gnp能很好地交联HbF,所得到的Gnp-HbF制品各项关键指标符合要求,为后续工艺降低了制备难度。Objective To achieve effective polymerization rate(molecular weight of 64-600 KD)≥70% and average molecular weight of 100-300 KD across the way of Gennipin(Genipin,Gnp)crosslinked hemoglobin isolated from fetal(hemoglobin isolated from fetal,HbF).Methods HbF was used as raw material,and Gnp as crosslinker.Orthogonal test involving four factors as hemoglobin concentration(HbC),molar ratio of Gnp to HbF,reaction time and reaction pH,and three levels were selected(n=3)to determine better matching condition.Three batches of Gnp-HbF samples were prepared under the optimal conditions,and the effective polymerization rate,dimer,average molecular weight,the content of methemoglobin(MetHb)and P_(50)were determined during the preparation process.Results The optimal matching conditions for Gnp-HbF obtained by orthogonal test were HbC:80g/L,molar ratio of Gnp to HbF:7∶1,reaction time:18 h,pH:8.5,with effective polymerization rate at(73.03±2.03)%,dimer(26.97±2.03)%,average molecular weight(167.70±28.40)KD,the content of methemoglobin(2.60±0.17)%and P_(50)(13.13±1.02)mmHg.Conclusion Gnp can effectively cross-link HbF,and the obtained key indicators of Gnp-HbF products meet the requirements,which reduces the preparation difficulty for the subsequent process.

关 键 词：京尼平(Gnp) 血红蛋白 正交试验/四因素 血红蛋白类氧载体 

分 类 号：R331.141[医药卫生—人体生理学] R714.51[医药卫生—基础医学]