title: Supercoiled circular DNA and protein retention in non-equilibrium chromatography temperature and velocity dependence: testimony of a transition.
abstract: Non-equilibrium chromatography (NEC) is a chromatographic mode for the rapid separation of polymers. The retention behavior of various proteins (human, chicken, bovine serum albumin) and supercoiled circular double-stranded DNA (plasmids) was investigated using a phosphate buffer as a mobile phase at different velocities and column temperatures with a C1 column with very low-packing particle diameter as a stationary phase. It was shown that the two factors (temperature and velocity) constituted important parameters in the retention mechanism of plasmids and proteins in NEC. The protein was retained more than the plasmid. At all the temperatures (5, 10, 15, 20, 25 degrees C) the plasmid retention increased over the entire flow-rate range (0.02-1.8 ml/min). For the protein, the retention curve presented a decrease in the relative retention time until a critical value of the mobile phase flow-rate, followed by an increase. The transition between the two well known NEC methods, slalom chromatography and hydrodynamic chromatography was clearly visualized for proteins at the lowest temperature, but did not appear for plasmids due to their strong compact structure.
authors: Laboratoire de Chemie Appliquée, La Valette du Var, France.
authors: François Xavier
authors: FX
authors: Perrin
authors: Carol Courderot
authors: CC
authors: Masuyer
authors: Tong-Thanh
authors: TT
authors: Truong
authors: Christiane
authors: C
authors: Guinchard
authors: Joëlle
authors: J
authors: Millet
authors: Jean Pierre
authors: JP
authors: Chaumont
authors: Mireille
authors: M
authors: Thomassin
authors: Yves Claude
authors: YC
authors: Guillaum
authors: Laurence
authors: L
authors: Nicod
mesh_terms: D002845:Chromatography / Q000379:methods*; D004278:DNA, Superhelical / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D013696:Temperature
chemical_list: D004278:DNA, Superhelical; D011506:Proteins