The importance of column design for protein separation using aqueous two-phase systems on J-type countercurrent chromatography

Guan, Yue; Bourton, Emma C.; Hewitson, Peter; Sutherland, Ian; Fisher, Derek

doi:10.1016/j.seppur.2008.07.016

Cited by 25 publications

(23 citation statements)

References 22 publications

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“…In addition, the length of the present 5-mm column is similar to the 25.8-m length "short" column for our 1.6-mm bore size column [6]. Conclusively, there has been no compromise of protein separation resolution after scaling up from the narrow to the present large bore size toroidal column.…”

Section: Separation On Model Proteins Using the 5-mm Bore Size Toroidsupporting

confidence: 53%

“…For mobile phase flow rates of 0.62, 1.25, 2.5 ml/min for our previous 1.6-mm narrow bore toroidal column [6], the corresponding flow rates for the present 5-mm wide bore column are 6.1, 12.3, 30.6 ml/min respectively.…”

Section: Resultsmentioning

confidence: 80%

“…This ATPS system is often shortened as "12.5% ATPS" in this paper. The physical properties of 12.5% ATPS system are: settling time 130 s, density difference 48.8 kg/m 3 , interfacial tension 0.095 mN m -1 , upper phase viscosity 4.84 mPa s, and low phase viscosity 1.71 mPa s [6,7]. The partition coefficients for the proteins or the dipeptides (see below) were measured and have been reported in Table 1 of [6].…”

Section: Phase System Preparationmentioning

confidence: 99%

“…Protein separation was made using the myoglobin and lysozyme protein pair [6,7,27,28] and initiated by injecting either myoglobin or lysozyme but not both. By plotting the resulting chromatograms on the same time abscissa, the potential is revealing in separating the two proteins at 10 ml/min lower mobile phase flow when stationary phase retention is at ca.…”

Section: Separation On Model Proteins Using the 5-mm Bore Size Toroidmentioning

confidence: 99%

“…ATPS stationary phase, the column geometry mounted on the CCC machinery needs to meet certain requirements [6,12,25]. Briefly, when the two immiscible phases (either two aqueous or two organic phases) have similar hydrophilicity or hydrophobicity, mobile phase flow tends to drag out the intended stationary phase and thus may considerably reduce stationary phase retention [12].…”

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confidence: 99%

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Scale-up protein separation on stainless steel wide bore toroidal columns in the type-J counter-current chromatography

Guan

Hewitson

Heuvel

et al. 2015

Journal of Chromatography A

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Manufacturing high-value added biotech biopharmaceutical products (e.g. therapeutic proteins) requires quick-to-develop, GMP-compliant, easy-to-scale and cost effective preparatory chromatography technologies. In this work, we describe the construction and testing of a set of 5-mm inner diameter stainless steel toroidal columns for use on commercially available preparatory scale synchronous J-type counter-current chromatography (CCC) machinery. We used a 20.2 m long column with an aqueous two-phase system containing 14% (w/w) PEG1000 and 14% (w/w) potassium phosphate at pH 7, and tested a sample loading of 5% column volume and a mobile phase flow rate of 20 ml/min. We then satisfactorily demonstrated the potential for a weekly protein separation and preparation throughput of ca. 11 g based on a normal weekly routine for separating a pair of model proteins by making five stacked injections on a single portion of stationary phase with no stripping. Compared to our previous 1.6 mm bore PTFE toroidal column, the present columns enlarged the nominal column processing throughput by nearly 10. For an ideal model protein injection modality, we observed a scaling up factor of at least 21. The 2 scales of protein separation and purification steps were realized on the same commercial CCC device.

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Section: Separation On Model Proteins Using the 5-mm Bore Size Toroidsupporting

confidence: 53%

Section: Resultsmentioning

confidence: 80%

Section: Phase System Preparationmentioning

confidence: 99%

Section: Separation On Model Proteins Using the 5-mm Bore Size Toroidmentioning

confidence: 99%

mentioning

confidence: 99%

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Scale-up protein separation on stainless steel wide bore toroidal columns in the type-J counter-current chromatography

Guan

Hewitson

Heuvel

et al. 2015

Journal of Chromatography A

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Studies on the performance of different coiled column configurations for compact type‐I countercurrent chromatography

Yang

Aisa

et al. 2011

J of Separation Science

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Three types of novel coiled column configurations, i.e., a triangular coiled column and elliptical coiled columns I and II, were designed for type-I countercurrent chromatography and their performances were evaluated with two solvent systems each with suitable test samples. Three DNP-amino acids (DNP-DL-glu, DNP-β-ala and DNP-L-ala) were separated with a moderately hydrophobic two-phase solvent system composed of hexane-ethyl acetate-metanol-0.1M hydrochloric acid (1:1:1:1, v/v), while two dipeptides (tryptophyl tyrosine and valyl-tyrosine) were separated with a polar solvent system composed of 1-butanol-acetic acid-water (4.75:0.25:5, v/v). The overall results indicated that the performance of compact type-I counter-current chromatography was improved by elliptical coiled column II which was mounted with its maximum coil diameter perpendicular to the surface of the column holder. Hydrodynamic effects involved in these separations were discussed.

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A simple gradient equilibrium method for better separation in countercurrent chromatography

Quan

Huang

et al. 2018

J of Separation Science

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The stationary phase retention is one of the most important parameters in countercurrent chromatography. In this work, a simple gradient equilibrium method was developed to further improve the stationary phase retention based on the optimized condition in the traditional equilibrium model. Meanwhile, this novel gradient equilibrium method was used to separate three flavone model compounds and compared with the conventional isocratic equilibrium method to evaluate the separation efficiency. The results show that better resolution or shorter separation time could be achieved with gradient equilibrium compared to isocratic equilibrium. So this novel equilibrium method has enormous potential for obtaining a better separation or saving the separating time in the preparative separation of target compounds.

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The importance of column design for protein separation using aqueous two-phase systems on J-type countercurrent chromatography

Cited by 25 publications

References 22 publications

Scale-up protein separation on stainless steel wide bore toroidal columns in the type-J counter-current chromatography

Scale-up protein separation on stainless steel wide bore toroidal columns in the type-J counter-current chromatography

Studies on the performance of different coiled column configurations for compact type‐I countercurrent chromatography

A simple gradient equilibrium method for better separation in countercurrent chromatography

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