新型人血白蛋白分离介质吸附性能及重组人血白蛋白分离  

Adsorption performance of a new resin for human serum albumin and recombinant human serum albumin purification

作　　者：葛程童 褚文宁 姚善泾[1] 林东强[1] GE Cheng-tong;CHU Wen-ning;YAO Shan-jing;LIN Dong-qiang(Key Laboratory of Biomass Chemical Engineering of Ministry of Education,College of Chemical and Biological Engineering,Zhejiang University,Hangzhou 310027,China)

机构地区：[1]浙江大学生物质化工教育部重点实验室,浙江大学化学工程与生物工程学院,浙江杭州310027

出　　处：《高校化学工程学报》2021年第3期492-499,共8页Journal of Chemical Engineering of Chinese Universities

基　　金：国家自然科学基金(21476198,21776243);国际科技合作专项(2015DFG42070)。

摘　　要：针对毕赤酵母发酵液中分离重组人血白蛋白(rHSA),采用新型介质IAA-CYS,探讨HSA吸附性能,优化分离条件,实现rHSA高效分离。考察了不同pH和盐浓度下IAA-CYS介质对HSA的吸附,发现pH为4.5是最佳条件,HSA饱和吸附容量达到157.6 mg×mL^(-1),盐浓度影响较小。测定了不同NaCl浓度和线性流速下IAA-CYS介质对HSA的动态载量,发现流速影响较大,合适流速为100 cm×h^(-1);盐浓度影响较小,无需对酵母发酵液进行稀释预处理,rHSA动态载量达48.3 mg×mL^(-1)。进一步优化了发酵液中分离rHSA,上样pH为4.5,上样流速为100 cm×h^(-1),上样量为38.6 mg×mL^(-1),在pH为5.0和质量浓度为400 mmol·L^(-1)的NaCl条件下淋洗,在pH为8.0和质量浓度为200 mmol·L^(-1)的NaCl条件下洗脱,rHSA纯度为90.1%,收率为88.6%。结果表明,IAA-CYS介质分离rHSA条件温和,耐盐性好,料液无需稀释预处理,过程简便高效,具有良好的应用前景。In order to separate recombinant human serum albumin(rHSA)from Pichia pastoris fermentation broth,HSA adsorption properties of a new resin IAA-CYS were studied and rHSA was separated from yeast broth after optimization.The adsorption of HSA with the IAA-CYS resin at different pH and salt concentrations was investigated.The results show that pH 4.5 is the best adsorption condition.Saturated adsorption capacity of HSA reached 157.6 mg×mL^(-1)with little salt effects.The dynamic binding capacities of HSA under different NaCl concentrations and linear velocities were determined.The results indicate that velocity has great effect,and the suitable linear velocity is 100 cm×h^(-1).The salt concentration had little influence,and yeast broth could be directly loaded without dilution.The dynamic binding capacity of rHSA reached 48.3 mg×mL^(-1).The separation of rHSA from yeast broth was further optimized and optimum loading conditions were pH 4.5,velocity 100 cm×h^(-1)and loading amount 38.6 mg×mL^(-1)resin.After washing with 400 mmol×L^(-1)NaCl at pH 5.0,rHSA was eluted with 200 mmol×L^(-1)NaCl at pH 8.0.The purity of rHSA reached 90.1%while the yield was 88.6%.The results indicate that the separation conditions for rHSA separation with IAA-CYS resin are mild,and the fermentation broth does not need dilution due to good salt tolerance.The process developed is simple and efficient,which indicates that IAA-CYS has good application potential for rHSA separation.

关 键 词：人血白蛋白 层析介质 吸附性能 蛋白分离 

分 类 号：Q819[生物学—生物工程]