Merck + Co., Inc.

Sangeetha Sagar

Ann L Lee

A process is disclosed for the large scale isolation and purification of plasmid DNA from large scale microbial fermentations. The process exploits a rapid heating method to induce cell lysis and precipitate genomic DNA, proteins and other debris while keeping the plasmid in solution. Suspending the microbial cells in buffer and then heating the suspension to about 70-100&deg; C. in a flow-through heat exchanger results in excellent lysis. Continuous flow or batch-wise centrifugation of the lysate effects a pellet that contains the cell debris, protein and most of the genomic DNA while the plasmid remains in the supernatant. This invention offers a number of advantages including higher product recovery than by chemical lyses, inactivation of Dnases, operational simplicity and scaleability.

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Lansdale, PA, United States of America

Sangeetha Sagar