Protein adsorption and transport in dextran-modified ion-exchange media. I: adsorption.
Adsorption behavior is compared on a traditional agarose-based ion-exchange resin and on two dextran-modified resins, using three proteins to examine the effect of protein size. The latter resins typically exhibit higher static capacities at low ionic strengths and electron microscopy provides direct visual evidence supporting the view that the higher static capacities are due to the larger available binding volume afforded by the dextran. However, isocratic retention experiments reveal that the larger proteins can be almost completely excluded from the dextran layer at high ionic strengths, potentially leading to significant losses in static capacity at relevant column loading conditions. Knowledge of resin and protein properties is used to estimate physical limits on the static capacities of the resins in order to provide a meaningful interpretation of the observed static capacities. Results of such estimates are consistent with the expectation that available surface area is limiting for traditional resins. In dextran-modified media, however, the volume of the dextran layer appears to limit adsorption when the protein charge is low relative to the resin charge, but the protein-resin electroneutrality may be limiting when the protein charge is relatively high. Such analyses may prove useful for semiquantitative prediction of maximum static capacities and selection of operating conditions when combined with protein transport information.
Bowes
Brian D
BD
Department of Chemical Engineering, University of Delaware, Newark, DE 19716, USA.
Koku
Harun
H
Czymmek
Kirk J
KJ
Lenhoff
Abraham M
AM
D000327:Adsorption; D000818:Animals; D002417:Cattle; D002645:Chickens; D002852:Chromatography, Ion Exchange / Q000295:instrumentation* / Q000379:methods; D003911:Dextrans / Q000737:chemistry*; D007475:Ion Exchange Resins / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D012117:Resins, Synthetic / Q000737:chemistry*
D003911:Dextrans; D007475:Ion Exchange Resins; D011506:Proteins; D012117:Resins, Synthetic