Supercoiled circular DNA retention in nonequilibrium chromatography: viscosity and velocity dependence--behavior difference with proteins.
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.
Equipe des Sciences Séparatives et Biopharmaceutiques (2SB), Laboratoire de Chimie Analytique, Faculté de Médecine et de Pharmacie, Place Saint Jacques, 25030 Besançon, Cedex, France. yves.guillaume@univ-fcomte.fr
Yves Claude
YC
Guillaume
Claire
C
André
Myriam
M
Matoga
Tong-Thanh
TT
Truong
Jean Louis
JL
Mozer
Mireille
M
Thomassin
Catherine
C
Guyon-Benay
Laurence
L
Nicod
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004278:DNA, Superhelical / Q000737:chemistry*; D010957:Plasmids
D004278:DNA, Superhelical