一种转基因猪血中重组人血清白蛋白分离纯化新方法
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摘要
基因工程技术已经成为研究和生产重组人血清白蛋白(rHSA)替代人血清白蛋白(HSA)的重点技术,而白蛋白的纯化则是该技术的关键。本文主要介绍了从转基因猪血中纯化rHSA的一种新方法,即热乙醇沉淀与多级色谱分离相结合的rHSA纯化方法。热乙醇沉淀法可从猪血浆中获得rHSA粗提取液,此时rHSA的纯度可达69.5%,回收率达51.3%。进一步采用多级色谱分离法,即阴离子交换色谱和反相色谱法进一步纯化,得到rHSA的最终纯度约为100.0%,总回收率为41.1%。该方法为从转基因猪血浆中大规模纯化用于临床和生化研究的高纯度rHSA提供可能,同时也为rHSA替代HSA奠定了基础。
Gene engineering technology has become a key technique for research and production of recombinant human serum albumin(rHSA) replacing human serum albumin(HSA),while the methods for rHSA purification are the key point to this technique.A novel method for rHSA purification from transgenic pig plasma was introduced in this paper,in which thermal ethanol precipitation combined with multi-stage liquid chromatography was chosen for rHSA purification.The rHSA was firstly extracted from pig plasma by the modified thermal ethanol precipitation method,with its purity and recovery of 69.5% and 51.3%,respectively.Then,the crude extract was further purified by multi-stage liquid chromatographic techniques,i.e.anion exchange chromatography and reversed-phase chromatography,with the purity and recovery for rHSA actually up to 100.0% and 41.1%,respectively.This method provides an opportunity for the large-scale purification of rHSA from transgenic pig plasma for clinical and biochemical research,and lays a foundation for the application of rHSA in place of HSA.
Gene engineering technology has become a key technique for research and production of recombinant human serum albumin(rHSA) replacing human serum albumin(HSA),while the methods for rHSA purification are the key point to this technique.A novel method for rHSA purification from transgenic pig plasma was introduced in this paper,in which thermal ethanol precipitation combined with multi-stage liquid chromatography was chosen for rHSA purification.The rHSA was firstly extracted from pig plasma by the modified thermal ethanol precipitation method,with its purity and recovery of 69.5% and 51.3%,respectively.Then,the crude extract was further purified by multi-stage liquid chromatographic techniques,i.e.anion exchange chromatography and reversed-phase chromatography,with the purity and recovery for rHSA actually up to 100.0% and 41.1%,respectively.This method provides an opportunity for the large-scale purification of rHSA from transgenic pig plasma for clinical and biochemical research,and lays a foundation for the application of rHSA in place of HSA.
引文
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[11] Pu Y Y.Kao Shi Zhou Kan(浦奕奕.考试周刊),2008,(3):236-237.
[12] Jiang H,Han W,Liu Y H,Zeng D Q,Li M,Zhao Y,Ma J.Med.J.Natl.Defend.Forces Southwest China(姜恒,韩威,刘彦华,曾德权,黎敏,赵燕,马骄.西南国防医药),2011,21(11):1167-1169.
[13] Li P,Yuan Z M,Tan Y Q.China Patent(李攀,袁正明,谈宇清.中国专利),CN 102952187A,2013.
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[15] Li Y,Yao J,Yang P Y,Fan H Z.Chem.J.Chin.Univ.(李颖,姚鋆,杨芃原,樊惠芝.高等学校化学学报),2015,36(6):1074-1081.