Ge Healthcare Bio-sciences Ab

Amersham Biosciences Ab

Joachim Stadler

James Van Alstine

Jan Berglof

Raf Lemmens

Dag Lindström

Tomas Nyhammar

Jan Bergström

Jan Amersham Biosciences Bergström

Dag Amersham Biosciences Lindström

James Amersham Biosciences Ab Van Alstine

The present invention relates to a method of isolating at least one plasmid from other components of a liquid, which method comprises the steps of providing a separation matrix comprised of one or more porous carriers, which carrier(s) present anion exchange groups on external surfaces as well as pore surfaces and a pore size distribution that does not allow access of plasmids to pore surfaces; contacting said matrix with the liquid to allow adsorption of the plasmids to ligands present on the separation matrix; contacting an eluent with the separation matrix to release the plasmids and recovering plasmids from a fraction of said eluent. Thus, the present method allows the plasmids to adsorb to the external surfaces of the matrix, while other components such as RNA is adsorbed onto the pore surfaces. In one embodiment, the matrix presents a DNA exclusion limit of at least about 270 base pairs; such as at least about 1,000 base pairs.

Plasmid purification

The present invention relates to a process for separating nucleic acid molecules, preferably open circular and supercoiled plasmid DNA and RNA molecules from each other, comprising the steps of providing a solution comprising the molecules; adsorbing the molecules to adsorbing groups on a carrier; and optionally washing the column with a suitable solution. The present process is especially suitable for large-scale isolation of supercoiled ccc DNA to be used in gene therapy.

Isolation of DNA molecules

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Freiburg, Germany

Joachim Stadler