A Method with Increased Yield for Production of Polysaccharide-Protein Conjugate Vaccines Using Hydrazide Chemistry
Current methods for synthesis and manufacturing of polysaccharide-protein conjugate vaccines employ conjugation reactions with low efficiency (about twenty percent). This means that up to eighty percent of the added activated polysaccharide (PS) is lost. In addition, inclusion of a chromatographic process for purification of the conjugates from unconjugated PS is required.
The present invention utilizes the characteristic chemical property of hydrazide groups on one reactant to react with aldehyde groups or cyanate esters on the other reactant with an improved conjugate yield of at least sixty percent. With this conjugation efficiency the leftover unconjugated protein and polysaccharide would not need to be removed and thus the purification process of the conjugate product can be limited to diafiltration to remove the by-products of small molecules. The new conjugation reaction can be carried out within one or two days with reactant concentrations between 1 and 25 mg/mL at PS/protein ratios from 1:2 to 3:1, at temperatures between 4 and 40 degrees Centigrade, and in a pH range of 5.5 to 7.4, optimal conditions varying from PS to PS.
Potential Commercial Applications:
Cost effective and efficient manufacturing of conjugate vaccines
Che-Hung Robert Lee
U.S. Pat: No. 8,048,432 issued 2011-11-01
U.S. Pat: No. 8,465,749 issued 2013-06-18
U.S. Pat: No. 8,753,649 issued 2014-06-17
PCT Application No. PCT/US2004/25477
PCT Application No. PCT/US2004/26431
US Application No. 10/566,898
US Application No. 14/270,213
International rights available
Bill Ronnenberg, JD-MIP, MS
FDA Technology Transfer Program
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OTT Reference No: E-301-2003/0
Updated: August 9, 2015