Electrostatic contributions to protein retention in ion-exchange chromatography. 2. Proteins with various degrees of structural differences.
The relation of protein structure to retention provides a framework within which to investigate chromatographic adsorption mechanisms. Protein sets with varying degrees of structural differences were studied to relate variations in protein properties to retention behavior. To explore molecular contributions to protein adsorption in ion-exchange chromatography, protein-adsorbent electrostatic interactions were modeled using a continuum approach. The calculations qualitatively capture the chromatographic differentiation of closely related subtilisin variants. Descriptions of the electrostatic interactions of FGF-1 vs FGF-2 with cation exchangers were obtained, and aid in rationalizing differences in experimental retention trends across a set of adsorbents based on different adsorption mechanisms linked to the adsorbent structure. Comparative calculations for proteins with differences in local or overall arginine-lysine composition, including subtilisin variants G166R/G166K and lysozyme/cytochrome c, suggest that continuum electrostatics is not adequate to capture the full quantitative characteristics of the chromatographic retention of proteins. To allow more accurate description of retention, additional molecular interactions, specifically hydration effects, must be incorporated in the model.
Yao
Yan
Y
Department of Chemical Engineering, University of Delaware, Newark, Delaware 19716, USA.
Lenhoff
Abraham M
AM
D001120:Arginine / Q000737:chemistry; D002412:Cations; D002852:Chromatography, Ion Exchange / Q000379:methods*; D045304:Cytochromes c / Q000737:chemistry / Q000235:genetics; D016220:Fibroblast Growth Factor 1 / Q000737:chemistry; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry; D008239:Lysine / Q000737:chemistry; D008956:Models, Chemical; D008958:Models, Molecular; D009113:Muramidase / Q000737:chemistry; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification; D055672:Static Electricity; D020860:Subtilisin / Q000737:chemistry
D002412:Cations; D011506:Proteins; D016222:Fibroblast Growth Factor 2; D016220:Fibroblast Growth Factor 1; D045304:Cytochromes c; D001120:Arginine; D009113:Muramidase; D020860:Subtilisin; D008239:Lysine