Inventors:
Yonghui Yuan - Tappan NY, US
Mark Ruppen - Garnerville NY, US
Wei-Qiang Sun - Morristown NJ, US
Ling Chu - Suffern NY, US
John Simpson - Upper Nyack NY, US
James Patch - Cornwall on Hudson NY, US
Justin Keith Moran - Valley Cottage NY, US
Pamela Fink - New City NY, US
International Classification:
C12P 19/04
Abstract:
A shortened process for producing a solution containing substantially purified capsular polysaccharides from a cellular lysate broth is described. Ultrafiltering and diafiltering a clarified lysate 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 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, the cells 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).