Wyeth

James Patch

Mark Edward Ruppen

Justin Keith Moran

John W Simpson

Wei-Qiang Willie Sun

Ling Chu

Yonghui Yuan

Pamela S Fink

Pamela Fink Charbonneau

Mark Ruppen

John Simpson

Justin K Moran

A shortened process for producing a solution containing substantially purified capsular polysaccharides from a cellularlysate broth is described. Ultrafiltering and diafiltering a clarifiedlysate followed by pH adjustment to less than 4.5, preferably about 3.5, precipitated at least 98% of the protein in the solution without seriously affecting polysaccharide yield. Furthermore, following ultrafiltration and diafiltration and acidification to a pH of less than 4.5, filtration using activated carbon precipitated at least 90% of remaining protein without seriously affecting polysaccharide yield. Exemplary, non-limiting S. pneumoniae serotypes that can be purified using the shortened process of the invention are 1, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F. In one embodiment, thecells are lysed using deoxycholate sodium (DOC), while in another embodiment the lytic agent is a non-animal derived lytic agent such as N-lauryl sarcosine sodium (NLS).

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A shortened process for producing a solution containing substantially purified capsular polysaccharides from a cellularlysate broth is described. Ultrafiltering and diafiltering a clarifiedlysate followed by pH adjustment to less than 4.5, preferably about 3.5, precipitated at least 98% of the protein in the solution without seriously affecting polysaccharide yield. Furthermore, following ultrafiltration and diafiltration and acidification to a pH of less than 4.5, filtration using activated carbon precipitated at least 90% of remaining protein without seriously affecting polysaccharide yield. Exemplary, non-limitingserotypes that can be purified using the shortened process of the invention are 1, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F. In one embodiment, thecells are lysed using deoxycholate sodium (DOC), while in another embodiment the lytic agent is a non-animal derived lytic agent such as N-lauryl sarcosine sodium (NLS).

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Cornwall on Hudson, NY, United States of America

James Patch