Merck + Co., Inc.

Ann L Lee

Robert D Sitrin

Mark S Rienstra

Walter E Manger

Chi Shung Brian To

Sangeetha Sagar

John O Konz, Jr

Aaron R Goerke

A process for purifying virus particles, especially recombinant adenovirus vector particles, is presented. The process relies on various combinations of cell lysis, detergent-based precipitation of host cell contaminants away from the virus, depth filtration or centrifugation, ultrafiltration, nuclease digestion and chromatography to robustly and economically produce highly purified product. This process results in contaminating DNA levels which are consistently below detectable levels.

Methods of adenovirus purification

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.

Method for large scale plasmid purification

A process for converting lipid-containing bacterial capsular polysaccharide, such as lipo-polyribosyl ribitol phosphate, lipo-PRP, into lipid-free, endotoxin-free polysaccharide, such as polyribosyl ribitol phosphate, PRP, by solubilizing polysaccharide-containing powder derived from culture media of bacteria, such as Haemophilus influenzae type b, cleaving covalently bound fatty acids from the polysaccharide, and removing the lipids, and endotoxin.

Process for converting lipid-containing bacterial capsular

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Lansdale, PA, United States of America

Ann L Lee