Protein folding liquid chromatography.
A method for carrying out protein folding with simultaneous separation by protein folding liquid chromatography (PFLC) is described herein. Furthermore, a two-dimensional chromatographic column, termed a 2D column, which can be independently employed for accomplishing PFLC in either weak cation exchange mode or hydrophobic interaction chromatography mode is reported. The content of this chapter describes the most commonly employed methods and operations of PFLC, such as the use of urea or guanidine hydrochloride as a denaturant with the protein in either the reduced or oxidized state and solving problems caused by the formation of the precipitates during protein folding. The PFLC can be performed using conventional chromatographic columns and a new chromatographic cake. A protocol for fast renaturation with simultaneous purification of inclusion body protein of the recombinant human interferon-gamma to obtain purity ≥95% and high specific bioactivity in a single step and in 1 h is introduced.
Key Laboratory of Modern Separation Science in Shaanxi Province, Key Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education, Institute of Modern Separation Science, Northwest University, Xi'an, China. baiquan@nwu.edu.cn
Quan
Q
Bai
Xindu
X
Geng
D002853:Chromatography, Liquid / Q000379:methods; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D017510:Protein Folding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*
D011994:Recombinant Proteins; D007371:Interferon-gamma