用于变性蛋白质复性及同时纯化的简易型色谱柱的研制及其应用

研制出一种简易型色谱柱并通过装填大颗粒的疏水色谱填料对色谱柱的性能进行了考察,该柱的外形和操作如同传统的液相色谱柱,但它却在生物大分子分离方面有着和高效液相色谱相似的分辨率。另外,只要给该柱装填合适的固定相,比如疏水相互作用色谱固定相,便可用于蛋白质的复性及同时纯化。实验考察了该色谱柱的结构、操作和性能,包括柱压、柱寿命及对蛋白质的分辨率等。以溶菌酶为模型蛋白质,实验测得其在初始浓度为50.0 g/L时的质量回收率和活性回收率分别为(96.6±1.3)%和(101.1±6.0)%。这种简易型色谱柱价格低廉

Design and Resolution of a Simple-Type Column for Renaturation with Simultaneous Purification of Proteins and Its Application to Lysozyme Refolding

A new kind of column called simple-type chromatographic column using the packings of large particles, which resembles columns used in traditional low pressure liquid chromatography in both shape and operation but has resolution of biopolymers similar to high performance liquid chromatography, was designed and investigated. In addition, so long as a suitable stationary phase, such as hydrophobic interaction chromatographic stationary phase, is packed, the column can be employed for the purification with simultaneous renaturation of proteins. The structure, operation and characteristics of the designed column including pressure drop, column life, and resolution were investigated. Lysozyme was selected as the model protein for investigating the renaturation. The recoveries of mass and bioactivity were found to be (96.6 +/- 1.3)% and (101.1 +/- 6.0)% respectively when the original concentration of lysozyme was 50.0 g/L. The simple-type column is very cheap and easy to use, and can be applied to the purification of proteins under the condition of low pressure. Thus, it has great potential applications for the purification and/or renaturation of proteins in large scale, down-stream processing of therapeutic proteins in biotechnology.