CAJ | 학술논문

目的建立从血浆冷沉淀中同时制备因子Ⅷ(factorⅧ,FⅧ)、纤维结合蛋白(fibronectin,Fn)和纤维蛋白原(fibrinogen,Fg)的方法.方法用缓冲液溶解抽提冷沉淀,获得的冷沉淀抽提液通过2次DEAE离子交换层析和1次聚乙二醇沉淀进行分离纯化来同时制备FⅧ、Fn和Fg.采用FⅧ∶C活性测定试剂盒检测制备的FⅧ活性,并计算FⅧ比活和回收率.分别用双向和单向免疫扩散试验对制备的Fn和Fg进行定性和定量检测,并计算回收率.结果制备的FⅧ的平均活性为16.48 U/ml,平均比活为29.84 U/mg,回收率＞60％.制备的Fn和Fg的蛋白纯度均＞90％,回收率分别为＞80％和＞75％.结论采用建立的方法可从冷沉淀中同时制备FⅧ、Fn和Fg.
목적 건립종혈장랭침정중동시제비인자Ⅷ(factorⅧ,FⅧ)、섬유결합단백(fibronectin,Fn)화섬유단백원(fibrinogen,Fg)적방법.방법 용완충액용해추제랭침정,획득적랭침정추제액통과2차DEAE리자교환층석화1차취을이순침정진행분리순화래동시제비FⅧ、Fn화Fg.채용FⅧ∶C활성측정시제합검측제비적FⅧ활성,병계산FⅧ비활화회수솔.분별용쌍향화단향면역확산시험대제비적Fn화Fg진행정성화정량검측,병계산회수솔.결과 제비적FⅧ적평균활성위16.48 U/ml,평균비활위29.84 U/mg,회수솔＞60％.제비적Fn화Fg적단백순도균＞90％,회수솔분별위＞80％화＞75％.결론 채용건립적방법가종랭침정중동시제비FⅧ、Fn화Fg.
Objective To establish a method for simultaneous preparation of factor Ⅷ (FⅧ),fibronectin (Fn) and fibrinogen (Fg) from plasma cryoprecipitate.Methods The plasma cryoprecipitate was dissolved and extracted by buffer.The cryoprecipitate solution was separated and purified by twice DEAE ion exchange chromatography and once polyethylene glycol precipitation to prepare FⅧ,Fn and Fg simultaneously.FⅧ:C activity of FⅧ was detected by a detection kit.Specific activity and recovery of FⅧ were calculated.Qualities and quantities of Fn and Fg were detected respectively by double and single immunodiffusion tests.Recoveries of Fn and Fg were calculated.Results F Ⅷ:C activity,specific activity and recovery of the prepared FⅧ were 16.48 U/ml,29.84 U/mg and ＞60％,respectively.Protein purities of the prepared Fn and Fg were both ＞90％,and recoveries of Fn and Fg were ＞ 80％ and ＞ 75％ respectively.Conclusion FⅧ,Fn and Fg can be prepared simultaneously from plasma cryoprecipitate by the established method.