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.
Ke
Congyu
C
Institute of Modern Separation Science, Shaanxi Key Laboratory of Modern Separation Science, Northwest University, Xi'an 710069, China.
Bai
Quan
Q
Geng
Xindu
X
D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation* / Q000379:methods; D009113:Muramidase / Q000737:chemistry / Q000302:isolation & purification; D017510:Protein Folding; D020673:Protein Renaturation; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification
D011506:Proteins; D009113:Muramidase