Matrix-assisted refolding of oligomeric small heat-shock protein Hsp26

Franzmann, Titus M.

doi:10.1016/j.ijbiomac.2006.02.026

Cited by 4 publications

(2 citation statements)

References 27 publications

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“…The main advantage of the LC refolding method is that it not only prevents the unfolded protein molecules from aggregating with each other but also simultaneously purifies or partially purifies the protein during the chromatographic process; thus, it is called protein folding liquid chromatography (PFLC) [13,18]. Ion exchange chromatography (IEC) is a widely used chromatographic method for protein purification; it was reported that about 70% of protocol for protein purification involved IEC, and now, IEC has been becoming one of the most frequently used LC refolding methods and has been applied to many proteins with high yields [19][20][21][22][23][24][25][26][27][28].…”

Section: Introductionmentioning

confidence: 99%

Solubilization and Refolding with Simultaneous Purification of Recombinant Human Stem Cell Factor

Wang

Liu

Wang

et al. 2008

Appl Biochem Biotechnol

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Recombinant human stem cell factor (rhSCF) was solubilized and renatured from inclusion bodies expressed in Escherichia coli. The effect of both pH and urea on the solubilization of rhSCF inclusion bodies was investigated; the results indicate that the solubilization of rhSCF inclusion bodies was significantly influenced by the pH of the solution employed, and low concentration of urea can drastically improve the solubilization of rhSCF when solubilized by high pH solution. The solubilized rhSCF can be easily refolded with simultaneous purification by ion exchange chromatography (IEC), with a specific activity of 7.8 x 10(5) IU x mg(-1), a purity of 96.3%, and a mass recovery of 43.0%. The presented experimental results show that rhSCF solubilized by high pH solution containing low concentration of urea is easier to be renatured than that solubilized by high concentration of urea, and the IEC refolding method was more efficient than dilution refolding and dialysis refolding for rhSCF. It may have a great potential for large-scale production of rhSCF.

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Section: Introductionmentioning

confidence: 99%

Solubilization and Refolding with Simultaneous Purification of Recombinant Human Stem Cell Factor

Wang

Liu

Wang

et al. 2008

Appl Biochem Biotechnol

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“…Ion exchange chromatography (IEC) is a widely used chromatographic method for protein purification. It was reported that about 70% of protocols for protein purification involved IEC, and now IEC has become one of the most frequently used LC refolding methods, and has been applied to many proteins with high yields (Liu et al, , 2007Franzmann, 2006;Langenhof et al, 2005;Li et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

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

Wang

Geng

2007

Biomedical Chromatography

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Protein refolding is a key step for the production of recombinant proteins, especially at large scales, and usually their yields are very low. Application of liquid chromatography to protein refolding is an exciting step forward for this field. In this work, recombinant human granulocyte colony-stimulating factor (rhG-CSF) expressed in Escherichia coli was renatured with simultaneous purification by ion exchange chromatography (IEC) with a Q Sepharose FF column. Several chromatographic parameters affecting the refolding yield of the denatured/reduced rhG-CSF, such as the urea concentration, pH value, concentration and ratio of reduced/oxidized glutathione in the mobile phase, as well as the flow rate of the mobile phase, were investigated in detail and indicated that the urea concentration and the pH value were of great importance. At the optimal conditions, the renatured and purified rhG-CSF was found to have a specific bioactivity of 3.0 x 10(8) IU/mg, a purity of 96%, and a mass recovery of 49%. Compared with the usual dilution method, the IEC method developed here is more effective for rhG-CSF refolding in terms of specific bioactivity and mass recovery.

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