Behavior of the inadvertent pH transient formed by a salt gradient in the ion-exchange chromatography of proteins.
The inadvertent pH transient produced when a stepwise change in salt concentration is used as the eluent in ion-exchange chromatography was studied theoretically using a local-equilibrium theory and experimentally using both strong-base and weak-base anion-exchange column packings. The accuracy of the local-equilibrium theory was verified by comparing it to a full numerical solution of the governing partial differential equations obtained using the method of characteristics. The predictions from the local-equilibrium theory were observed to largely agree with experimental results. Detailed comparisons of experimental results and the local-equilibrium theory permitted the observed trends for the pH transients to be interpreted in terms of the physical properties of the column packing and mobile phase. The results of this study are useful for the design of ion-exchange processes using salt gradient elution where it is desired to limit the exposure of eluted proteins to the inadvertent pH transient caused by the salt gradient.
Pérez
Jessica Soto
JS
Department of Chemical and Biochemical Engineering, University of Maryland Baltimore County, Baltimore, Maryland 21250, USA.
Frey
Douglas D
DD
D002852:Chromatography, Ion Exchange / Q000379:methods*; D003198:Computer Simulation; D004058:Diffusion; D006863:Hydrogen-Ion Concentration; D008956:Models, Chemical; D011506:Proteins / Q000032:analysis* / Q000737:chemistry*; D012492:Salts / Q000737:chemistry*; D013997:Time Factors; D014325:Tromethamine / Q000737:chemistry
D011506:Proteins; D012492:Salts; D014325:Tromethamine