Theoretical background of monolithic short layer ion-exchange chromatography for separation of charged large biomolecules or bioparticles.
The retention and peak spreading in linear gradient elution of charged large biomolecules were investigated by using numerical simulations. Oligo-DNA separation by monolithic anion-exchange chromatography was chosen as a model system. The peak width and the retention were well predicted by using the parameters obtained by gradient elution experiments at different gradient slopes. As the distribution coefficient at the peak retention volume K(R) decreases with increasing molecular size, the peak became sharper for larger DNAs. This is due to very large effective charge (binding site) values of large DNAs (20-60). The peak width was well correlated with K(R) based on the model equation developed for linear gradient elution of proteins. It was shown that the monolithic disk is best suited for very large charged biomolecule separations at high flow velocities with shallow gradients slopes.
Yamamoto
Shuichi
S
Bio-Process Engineering Laboratory, School of Engineering and Graduate School of Medicine, Yamaguchi University, Tokiwadai, Ube 755-8611, Japan. shuichi@yamaguchi-u.ac.jp
Yoshimoto
Noriko
N
Nishizumi
Yukiko
Y
D002852:Chromatography, Ion Exchange / Q000379:methods*; D003198:Computer Simulation; D004247:DNA / Q000302:isolation & purification*; D008962:Models, Theoretical
D004247:DNA