重组人β淀粉样蛋白1-42小规模发酵及小量纯化工艺的摸索  

Small scale fermentation and a small purification technology of recombinant human β-amyloid peptide 1-42

作　　者：申茉函[1] 王全才[2] 杨丽[3] 

机构地区：[1]中国医科大学附属盛京医院,沈阳110004 [2]辽宁省人民医院,沈阳110016 [3]吉林大学人兽共患研究教育部重点实验室,长春130021

出　　处：《实用药物与临床》2015年第3期256-259,共4页Practical Pharmacy and Clinical Remedies

基　　金：国家高技术研究发展计划(863计划)(2004AA205020)

摘　　要：目的研究重组人β淀粉样蛋白1-42(rh Aβ1-42)在5 L摇瓶中的高密度发酵实验,以及小量纯化工艺的方法。方法分别从甲醇浓度、p H、诱导时间等方面对毕赤酵母重组菌株产生rh Aβ1-42的发酵过程进行了优化;通过硫酸铵沉淀目的蛋白、透析、脱盐和阴离子交换层析,对rh Aβ1-42小量精确的纯化方法进行了研究。结果确定rh Aβ1-42在毕赤酵母中分泌表达的最佳条件为:在p H 6.0的条件下,以0.5%甲醇诱导72 h,经过硫酸铵沉淀目的蛋白、透析、脱盐和阴离子交换层析,rh Aβ1-42纯度可达94%以上。结论确定了rh Aβ1-42在毕赤酵母中分泌表达的最佳条件,建立了小量纯化rh Aβ1-42的新方法。Objective To study the high-cell-density fermentation experiment of human recombinant β-amyloid protein 1-42（ rh Aβ1-42） in 5 L shake flask and the method of small scale purification. Methods Fermentation parameters of rh Aβ1-42 was further optimized mainly on concentration of methanol induction,pH value,and induced time. Secreted supernatant of rh Aβ1-42 was purified with ammonium sulphate sediment,dialysis desalination and anionexchange chromatography. Results The optimal condition of rh Aβ1-42 expressed in Pichia pastoris was： under the condition of pH = 6,induced by 0. 5% methanol for 72 h,precipitation of target protein by ammonium sulfate,dialysis desalting,anion exchange chromatography,rh Aβ1-42 purity could reach more than 94%. Conclusion A stable fermentation and new technology of the small scale purification is successfully set up.

关 键 词：Β淀粉样蛋白 发酵 小量纯化 

分 类 号：R749.16[医药卫生—神经病学与精神病学]