Purification of Capsular Polysaccharide Produced by Streptococcus pneumoniae Serotype 19A

Streptococcus pneumoniae is a major cause of invasive infection in young infants and older adults. There are currently 90 capsular serotypes identified and 23 serotypes (1, 2, 3, 4, 5, 68, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19F, 19A, 20, 22F, 23F, and 33F) are responsible for about 90% of invasive disease. Among the more than 90 different S. pneumoniae serotypes, serotype 19A is globally very prevalent. A simplified purification procedure including adjustment of cell lysate pH to 4.5, fractionation with 50-80% ethanol, and dialysis rendered capsular polysaccharide (CPS) in a yield of 31.32 +/- 3.11 mg from 1 1 culture (75% recovery after lyses). The product contained only 69.6 mu g of protein (99.78% purity) and 0.8 mg (sum of the precipitants from 50 similar to 60%, 60 similar to 70%, and 70 similar to 80%) of nucleic acid (97.45% purity). The purified CPS was conjugated with bovine serum albumin; the product size ranged from 100 to 180 kDa.