The thermodynamic principles of ligand binding in chromatography and biology.
In chromatography, macromolecules do not adsorb in the traditional sense of the word but bind to ligands that are covalently bonded to the surface of the porous bead. Therefore, the adsorption must be modelled as a process where protein molecules bind to the immobilised ligands. The paper discusses the general thermodynamic principles of ligand binding. Models of the multi-component adsorption in ion-exchange and hydrophobic chromatography, HIC and RPLC, are developed. The parameters in the models have a well-defined physical significance. The models are compared to the Langmuir model. In the traditional adsorption models, the standard state Gibbs energy change of adsorption does not depend level of occupancy, but when it depends on the level of occupancy it gives rise to an adsorptive behaviour known as cooperativity. The binding of oxygen to haemoglobin is a well-known example from biology but it is also observed in chromatography due to protein-protein interactions. Retention measurements on beta-lactoglobulin A demonstrate this. A discussion of salt effects on hydrophobic interactions in precipitation and chromatography of proteins concludes the paper.
Mollerup
Jørgen M
JM
Department of Chemical Engineering, Building 229, Technical University of Denmark, DK-2800 Lyngby, Denmark. jm@kt.dtu.dk
D000327:Adsorption; D002852:Chromatography, Ion Exchange; D057927:Hydrophobic and Hydrophilic Interactions; D008024:Ligands; D008954:Models, Biological; D011506:Proteins / Q000302:isolation & purification*; D013816:Thermodynamics
D008024:Ligands; D011506:Proteins