Evaluation of radial chromatography versus axial chromatography, practical approach.
Developments in packing and packing port design of radial columns in recent years have resulted in a claimed significant increase in performance of this process chromatography technology. In this first study, the main chromatographic parameters as efficiency, capacity factor, asymmetry and resolution were evaluated in a unique one-to-one comparison between a 120 ml bed-volume and 6 cm bed length radial chromatography mini-process column against a 50 mm diameter, 6 cm bed height and 120 ml bed-volume axial chromatography column. Radial chromatography showed an increase in efficiency by 31% in the number of plates per meter while the equilibration could be reduced by 0.4-0.5 column volumes. The asymmetry factor for bovine serum albumin in radial chromatography showed a reduction of 20% while the reduction of the asymmetry factor of the smaller protein ovotransferrin decreased even by 46% in comparison to the performance of the comparative axial chromatography column. Therefore in radial chromatography resolution improved up to 20%. The retention volume was similar in both cases. For radial chromatography, the decrease in "width at half height" at Height Equivalent of Theoretical Plates (HETP) measurements was 40% while the decrease of the over-all width of the peak was 27%. For adsorbed/desorbed proteins, the elution peak showed similar results: "width at half height" decreased to 45% while the over-all width of the peak decreased by 28%. The concentration of the non-retained protein in the flow-through (lysozyme), increased by 35% while the concentration of the eluted fraction (serum albumin bovine), increased with 40% in the radial chromatography columns. The better results obtained with the radial column were probably the consequence of the geometrical design of this device (larger inlet surface area and small outlet surface area which concentrate the eluted fraction).
Cabanne
Charlotte
C
ESTBB, Université Victor Segalen Bordeaux2, 146 rue Léo Saignat, 33076 Bordeaux cedex, France. charlotte.cabanne@estbb.u-bordeaux2.fr
Raedts
Marcel
M
Zavadzky
Emil
E
Santarelli
Xavier
X
D002853:Chromatography, Liquid / Q000295:instrumentation* / Q000379:methods; D003207:Conalbumin / Q000302:isolation & purification; D009113:Muramidase / Q000302:isolation & purification; D012710:Serum Albumin, Bovine / Q000302:isolation & purification
D003207:Conalbumin; D012710:Serum Albumin, Bovine; D009113:Muramidase