Supercoiled circular DNA retention in nonequilibrium chromatography: viscosity and velocity dependence--behavior difference with proteins |
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Authors: | Guillaume Yves Claude André Claire Matoga Myriam Truong Tong-Thanh Mozer Jean Louis Thomassin Mireille Guyon-Benay Catherine Nicod Laurence |
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Institution: | Equipe des Sciences Séparatives et Biopharmaceutiques (2SB), Laboratoire de Chimie Analytique, Faculté de Médecine et de Pharmacie, Place Saint Jacques, 25030 Besan?on, Cedex, France. yves.guillaume@univ-fcomte.fr |
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Abstract: | 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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