Published:Journal of Chromatographic Science,
ISSN 0021-9665Volume
39, Number 10, October 2001, pp. 133-136
Supercoiled Circular DNA Retention in Nonequilibrium Chromatography:
Viscosity and Velocity Dependence— Behavior Difference with Proteins
Yves Claude Guillaume[1], Claire André[1], Myriam Matoga[1],
Tong-Thanh-Truong[1], Jean Louis Mozer[3], Mireille Thomassin[1], Catherine
Guyon-Benay[1], and Laurence Nicod[2]
Equipe des Sciences Séparatives et Biopharmaceutiques (2SB), [1]Laboratoire
de Chimie Analytique, [2]Laboratoire de Biologie Cellulaire, [3]Laboratoire
de Biochimie, Faculté de Médecine et de Pharmacie, Place Saint
Jacques, 25030 Besançon, Cedex, France
This study demonstrates that
the retention behavior of various circular double-stranded DNA molecules (3,
5, and 10 kb) increases
over the entire flow-rate range (0.02–1.8 mL/min) at all the mobile phase
viscosities (h). The transition between the two well-known nonequilibrium chromatography
methods (slalom and hydrodynamic chromatography) is clearly visualized for proteins
and does not appear for plasmids because of their strong compact structure. Also,
the optimal conditions for F and h are determined to obtain the most efficient
separation of these three plasmids in a minimum analysis time.
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