Protein folding liquid chromatography and its recent developments

Abstract

The ultimate goal of proteomics is to identify biologically active proteins and to produce them using biotechnology tools such as bacterial hosts. However, proteins produced by Escherichia coli must be refolded to their native state. Protein folding liquid chromatography (PFLC) is a new method developed in recent years, and it is widely used in molecular biology and biotechnology. In this paper, the new method, PFLC is introduced and its recent development is reviewed. In addition the paper includes definitions, advantages, principles, applications for both laboratory and large scales, apparatus, and effecting factors of PFLC. In addition, the role of this method in the future is examined.

Renaturation with simultaneous purification of rhG‐CSF from Escherichia coli by ion exchange chromatography

“…Application of IEC to the refolding and simultaneous purification of rhG-CSF As mentioned above, LC can be a potential tool for protein refolding with high yield (Geng and Wang, 2007;Jungbauer et al, 2004;Li et al, 2004). So far, several LC methods, including affinity chromatography (AFC), ion exchange chromatography, hydrophobic interaction chromatography (HIC) and size exclusion chromatography (SEC) can be used for this purpose (Geng and Wang, 2007).…”

“…In recent years, liquid chromatography (LC) has been used to refold proteins with higher yields (Geng and Chang, 1992;Geng and Wang, 2007;Wang et al, 2004Wang et al, , 2006Jungbauer et al, 2004;Li et al, 2004). The advantages of the LC method are that it not only prevents the unfolded protein molecules from aggregating with each other, but it also simultaneously purifies or partially purifies the protein during the chromatographic process; thus it is called protein folding liquid chromatography (PFLC; Geng and Wang, 2007;Geng et al, 2006).…”

“…The advantages of the LC method are that it not only prevents the unfolded protein molecules from aggregating with each other, but it also simultaneously purifies or partially purifies the protein during the chromatographic process; thus it is called protein folding liquid chromatography (PFLC; Geng and Wang, 2007;Geng et al, 2006). Its recent developments and applications have been reviewed by several groups in different aspects (Geng and Wang, 2007;Jungbauer et al, 2004;Li et al, 2004). Ion exchange chromatography (IEC) is a widely used chromatographic method for protein purification.…”

Hydrophobic Interaction Chromatography: Fundamentals and Applications in Biomedical Engineering

“…Then, HIC often forms part of processes to yield a purified macromolecule of biomedical interest, such as therapeutic proteins (Seely & Richey, 2001), DNA vaccines (Diogo et al, 2000), and enzymes (Teng et al, 2010), among others. Besides, the use of HIC to purify protein complexes (McCue et al, 2008), as well as to study protein folding from a thermodynamic point of view (Geng & Wang, 2007), have been reported. Some applications of HIC for purifying enzymes and protein complexes, and to studying protein folding are described below.…”

“…At 25°C, ∆H folding was found to be -34 439 KJ/mol, involving adsorption, dehydration and molecular conformation enthalpies changes. Later, Geng & Wang (2007) used the concept of "Protein Folding Liquid Chromatography" (PFLC), to describe a chromatographic process aiming to either raise the efficiency, or shortening the time of protein folding. Besides, an optimal PFLC should be able to simultaneously remove denaturant substances, separate contaminant proteins, promote refolding of the target protein, and ease denaturant recovery.…”