Patent ID: 8895014

Claim:
A method of producing a composition comprising a bioconjugate in a bioreactor, wherein said bioconjugate comprises a protein linked to an O antigen, comprising the steps: (i) expressing in bacteria: modified EPA protein containing at least one consensus sequence, D/E-X-N-Z-S/T, wherein X and Z may be any natural amino acid except proline, or the AcrA protein; an oligosaccharyl transferase; and one or more O antigens; (ii) growing the bacteria for a period of time to produce an amount of the bioconjugate comprising the AcrA protein or the modified EPA protein linked to the one more O antigens; (iii) extracting periplasmic proteins; and (iv) separating the bioconjugate from the extracted periplasmic proteins by anionic exchange.