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AssigneeBoehringer Ingelheim Rcv GmbH & Co KG
InventorsJochen URTHALER, Christine ASCHER, Daniel BUCHELI
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A scalable process and device for producing a bio molecule, in particular pharmaceutical grade plasmid DNA is described. The process includes the steps of alkaline lysis, neutralization and clarification and can be further extended. For separating the lysate and the precipitate an improved floatation method is disclosed. This method is based on attachment of CO2 bubbles on the precipitate floe. The CO2 is released from a carbonate salt during or after neutralization (acidification). The method of the invention is preferably carried out in an automated continuous mode applying devices for lysis and neutralization and a novel device for completely continuous clarification (separation of flocs and clarified lysate).
A method for producing a biomolecule of interest that is not secreted by the host cells, comprising the steps of a) cultivating host cells to produce the biomolecule of interest and optionally harvesting and resuspending the cells, b) disintegrating the cells by alkaline lysis, c) neutralizing the lysate obtained in step b), whereby a precipitate is formed, d) separating the cleared lysate from the precipitate obtained in step c), e) purifying the biomolecule of interest, wherein a carbonate salt is added in at least one of step a-c), whereby due to acidic conditions at and after step c) CO2 is released and wherein in step d) the precipitate and the lysate are allowed to separate in a clarification device.
Dependencies are inferred from the claim text.