Conjugate purification

摘要

This application relates to methods for the purification of saccharide antigen-carrier protein conjugates. In particular, the invention provides a method for purifying saccharide antigen-carrier protein conjugates from free carrier protein, such as CRM1 97, using hydroxyapatite. The invention further relates to methods of preparing vaccines, using this method.

主权项

1. A method of separating capsular saccharide antigen conjugated to a carrier protein from the carrier protein not conjugated to the capsular saccharide antigen in a mixture comprising the carrier protein not conjugated to the capsular saccharide antigen and the capsular saccharide antigen conjugated to the carrier protein, wherein the capsular saccharide antigen conjugated to the carrier protein is from N. meningitidis, S. pneumoniae, S. pyogenes, S. agalactiae, H. influenzae, P. aeruginosa, S. aureus, E. faecalis, E. faecium, V. cholerae, or S. typhi, comprising
contacting said mixture with hydroxyapatite under conditions that allow the binding of the carrier protein not conjugated to the capsular saccharide antigen to the hydroxyapatite, and wherein the capsular saccharide antigen conjugated to the carrier protein does not bind to the hydroxyapatite; and collecting the capsular saccharide antigen conjugated to the carrier protein unbound to the hydroxyapatite.