Comparison of particulate and continuous-bed columns for protein displacement chromatography.
A conventional anion exchange column packed with porous particles (BioScale Q2), and a novel continuous-bed column (UNO Q1) were compared for displacement separation of dairy whey proteins with polyacrylic acid as displacer. The steric mass action model was investigated as a means to aid and accelerate this development. Characteristic charges and steric factors were measured for the proteins and the displacer according to the model, and used together with the affinity constant derived from the adsorption isotherms for simulations, as well as for the construction of the affinity and operating regime plots. If possible, the latter two were used to select conditions for the actual experiments. In the case of the particle-based column, experimental results and simulations did not agree. In addition, the operating regime plot could not be constructed. The affinity plot did predict the order in the displacement train correctly, but gave misleading information concerning the possible effect of a change in displacer concentration. This is taken to be a result of the porous nature of the particles, which handicaps, to some extent, the interaction of the proteins and the displacer molecules with the adsorptive surface. Results were considerably better in case of the continuous-bed column, where there is no intraparticulate surface.
Freitag
R
R
Department of Chemistry, Ecole Polytechnique Fédérale de Lausanne, Switzerland. ruth.freitag@epfl.ch
Vogt
S
S
D000180:Acrylic Resins; D000327:Adsorption; D000837:Anion Exchange Resins; D001709:Biotechnology / Q000379:methods; D002845:Chromatography / Q000295:instrumentation* / Q000379:methods*; D003612:Dairying; D007768:Lactalbumin / Q000032:analysis* / Q000302:isolation & purification*; D007782:Lactoglobulins / Q000032:analysis / Q000302:isolation & purification; D010316:Particle Size
D000180:Acrylic Resins; D000837:Anion Exchange Resins; D007782:Lactoglobulins; C006903:carbopol 940; D007768:Lactalbumin