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Amino Acids

, Volume 46, Issue 1, pp 153–165 | Cite as

Protein renaturation with simultaneous purification by protein folding liquid chromatography: recent developments

  • Lili Wang
  • Xindu GengEmail author
Invited Review

Abstract

Protein folding liquid chromatography (PFLC) is a powerful tool for protein refolding with simultaneous purification. We review its recent progress in liquid chromatography and molecular biology, primarily involving the validation of PFLC refolding of proteins containing multiple disulphide bonds, the application of mixed-mode chromatography, PFLC in molecular biology. Representative examples are described.

Keywords

Protein folding liquid chromatography (PFLC) Proteins Inclusion bodies Refolding with simultaneous purification Mixed-mode liquid chromatography 

Abbreviations

PFLC

Protein folding liquid chromatography

LC

Liquid chromatography

MMC

Mixed-mode chromatography

E. coli

Escherichia coli

N state

Native state

U state

Unfold state

M state

Intermediates state

HIC

Hydrophobic interaction chromatography

IEC

Ion exchange chromatography

SEC

Size exclusion chromatography

AFC

Affinity chromatography

RPLC

Reverse phase liquid chromatography

EBA

Expanded bed chromatography

SMB

Simulated moving bed

PFC

Perfusion chromatography

USRPP

The units of simultaneous renaturation and purification of proteins

AMC

Artificial molecular chaperone

AMC-IEC

AMC-ion exchange chromatography

GdnHCl

Guanidine hydrochloride

DTT

Dithiothreitol

β-ME

β-Mercaptoethanol

HI-EBAC

Hydrophobic interaction expanded bed adsorption chromatography

INAC

Immobilised nickel affinity chromatography

D-LC-1C

A single column

2D column

Two-dimensional column

2D-LC-1C

2D with a single column

WCX

Weak cation exchange

WAX

Weak anion-exchange chromatography

STHIC

Stationary phase of hydrophobic interaction chromatography

rhG-CSF

Recombinant human granulocyte colony stimulating factor

NCp-NiV

Nipah virus

HBx

Hepatitis B virus X

rhIFN-γ

Recombinant human interferon-γ

rhEGF

Recombinant human EGF

rhSCF

Recombinant human stem cell factor

rhAFP

Recombinant alpha-fetoprotein

rhFL

Recombinant human Flt3 ligand

rhIFN-alpha-2b

Recombinant human interferon-alpha 2b

LK68

Lipoprotein kringle

rhVEGF

Recombinant human vascular endothelial growth factor

rhDll1

Recombinant human Notch ligand delta-like 1

GSH/GSSG

Glutathione/glutathione

α-Chy

α-Chymotrypsin

Cyt C

Cytochrome C

MOP

Molecular orientation of protein

SDT

Stoichiometric displacement theory

PEG

Polyethylene glycol

CBD

Cellulose-binding domain

PPI

Peptidyl prolyl cis trans isomerase

PDI

Protein disulphide isomerase

α-Amy

α-Amylase

LC–MS

LC–mass spectrometry

Notes

Acknowledgments

This work was supported by the Foundation of Key Laboratory of Modern Separation Science in Shaanxi Province (Nos. 2010JS104; 11JS098), the Foundation of Science and Technology in Shaanxi Province (No. 2010K12-01-05), and the foundation of Provincial Key Description of Analytical Chemistry of Shaanxi Subject (ZDXKL00402).

Conflict of interest

The authors declare that they have no conflict of interest.

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© Springer-Verlag Wien 2013

Authors and Affiliations

  1. 1.Key Laboratory of Synthetic and Natural Functional Molecular Chemistry of Ministry of Education, Shaanxi Key Laboratory of Modern Separation Science, Institute of Modern Separation ScienceNorthwest UniversityXi’anChina

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