The present invention provides improved methods for the reduction or
removal of protein impurities from a complex cellular Streptococcus
pneumoniae lysate or centrate comprising serotype 3 polysaccharides
involving steps relating to post-lysis heating or pH adjustment. In
certain methods, the lysate is heated for a time and at a temperature
sufficient to denature proteins present in the lysate and cause their
aggregation and precipitation. In one embodiment, the lysate is heated to
at least 60.degree. C. for at least 30 minutes to cause protein
aggregation and precipitation, more particularly about 60.degree. C. to
about 70.degree. C. for about 30 to about 50 minutes, and even more
particularly about 65.degree. C. for about 40 minutes. In other methods,
the pH of the lysate or centrate is increased to at least 8.0 to improve
filterability, more particularly about 8.0 to 8.4, and even more
particularly about 8.2. In further methods, heating and pH adjustment
steps are combined to cause the aggregation and precipitation of proteins
as well as to improve filterability of the lysates or centrates. In other
methods, the pH of the lysate or centrate is lowered to about 3.0 to
about 5.0 to cause protein aggregation and precipitation. Such methods
allow for the production of substantially purified serotype 3
polysaccharide-containing lysates or centrates.