Abstract
Chromatography is used not only for high performance analysis but also for process scale purification of biomolecules. It is known that when a small difference in similar molecules is well recognized by chromatography stationary phase (ligand) a good separation performance can be obtained. Often intended and unintended conformational changes of biomolecules occur before or during chromatography. The former is pretreatment of DNA samples by heating for DNA chromatography. The latter includes denaturation of biomolecules due to adsorption. Other conformational changes during chromatography are refolding of (denatured) proteins which is called "solid-phase refolding" and PEGylation of proteins on chromatography (selective PEGylation and separation). In this study biorecognition and transport phenomena for during chromatography was investigated by using a model developed for ion-exchange chromatography. The samples employed are DNAs, denatured proteins and PEGylated proteins. The information on the number of bindings sites was carefully investigated along with the data obtained by separate experiments (Taylor method, melting-curve analysis, etc.).
