	US 11,883,502 B2
	*Methods for production of capsular polysaccharide protein conjugates from Streptococcus pneumoniae* serotype 19F**
Michael Albert Winters, Doylestown, PA (US); and John E. MacNair, Lansdale, PA (US)
Assigned to Merck Sharp & Dohme LLC, Rahway, NJ (US)
Filed by Merck Sharp & Dohme LLC, Rahway, NJ (US)
Filed on Dec. 7, 2021, as Appl. No. 17/544,239.
Application 17/544,239 is a continuation of application No. 16/482,337, abandoned, previously published as PCT/US2018/015905, filed on Jan. 30, 2018.
Claims priority of provisional application 62/452,521, filed on Jan. 31, 2017.
Prior Publication US 2022/0088210 A1, Mar. 24, 2022
Int. Cl. A61K 47/64 (2017.01); A61K 39/09 (2006.01); A61K 47/36 (2006.01); G01N 33/569 (2006.01); A61K 39/00 (2006.01)

CPC A61K 47/646 (2017.08) [A61K 39/092 (2013.01); A61K 47/36 (2013.01); G01N 33/56944 (2013.01); A61K 2039/6037 (2013.01); G01N 2333/315 (2013.01)]

8 Claims

1. A method for the production of a stable Streptococcus pneumoniae serotype 19F polysaccharide-CRM197 carrier protein conjugate, wherein the Streptococcus pneumoniae serotype 19F polysaccharide-CRM197 carrier protein conjugate comprises Streptococcus pneumoniae serotype 19F capsular polysaccharide covalently linked to a CRM197 carrier protein, the method comprising the steps of:

a) performing a conjugation reaction wherein Streptococcus pneumoniae serotype 19F polysaccharide is covalently linked to a CRM197 carrier protein, wherein the conjugation reaction generates a reaction mixture comprising Streptococcus pneumoniae serotype 19F polysaccharide-CRM197 carrier protein conjugates and free Streptococcus pneumoniae serotype 19F polysaccharides;

b) incubating said reaction mixture for 5-7 days at a temperature of 20-25° C. in a buffer at a pH of 6.5-7.5; and

c) performing size separation under conditions that allow removal of the free Streptococcus pneumoniae serotype 19F polysaccharides.