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.
Guillaume
Yves Claude
YC
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
André
Claire
C
Matoga
Myriam
M
Truong
Tong-Thanh
TT
Mozer
Jean Louis
JL
Thomassin
Mireille
M
Guyon-Benay
Catherine
C
Nicod
Laurence
L
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004278:DNA, Superhelical / Q000737:chemistry*; D010957:Plasmids
D004278:DNA, Superhelical