摘要
首次采用自制的分别级合了辛基、丁基、苯基及聚乙二醇-4000的4种常用疏水基团的纤维素疏水膜色谱柱，以键合了辛基、苯基的SepharoseCL－4B凝胶柱为对照，考察了疏水膜色谱柱对牛血清白蛋白（BSA）的动态吸附容量及流速对吸附容量的影响。疏水膜色谱柱对蛋白及酶具有较好的疏水吸附及纯化作用，但吸附容量比相应的琼脂糖凝胶柱低得多。增大流速及降低蛋白溶液质量浓度对疏水膜色谱柱的吸附容量影响较小，这些性能使膜色谱柱非常适合于大体积低质量浓度蛋白溶液（如基团工程培养液）的分离纯化。牛肝过氧化氢酶（BLC）经苯基膜色谱柱一步纯化，比活力提高11．8倍，蛋白回收率近100％，分离过程仅十几分钟。
Cellulose membrane bonded with four commonly used hydrophobic groups, octyl, butyl, phenyl and polyethylene glycol was first investigated for their binding and purification characteristics of protein and enzyme with octyl- and phenyl-Sepharose CL-4B as controls. Hydrophobic membranes bound BSA effectively by hydrophobic interaction in high salt solution. Their binding capacities were not notably affected by significantly increasing the flow mass rate or decreasing the mass concentration of protein solution, but were much lower than those of octyl- and phenyl-Sepharose CL-4B. 11. 8 fold of purification with an approximately 100% recovery of bovine liver catalase was achieved by step gradient elution on the phenyl cellulose membrane cartridge in a single step in only ten mins or a little more. Increase of the flow mass rate had no effecton the purification of catalase, however, the processing time was shortened greatly. Hydrophobic membrane chromatography here reported exibits a potential of fast processing of the protein solution in large volume with low mass concentration of the target protein, such as genetic engineering culture solution.
出处
《色谱》
CAS
CSCD
北大核心
1999年第4期335-338,共4页
Chinese Journal of Chromatography
基金
国家自然科学重点基金!29635010
关键词
膜色谱
分离
纯化
生物大分子
BSA
BLC
疏水色谱
membrane chromatography
purification
hydrophobic interaction
biological macromolecule
