Patent
In one aspect, the invention relates to a non-lipidated and non-pyruvylated Neisseria meningitidis serogroup B polypeptide and methods of use thereof. In another aspect, the invention relates to an immunogenic composition including an isolated non-lipidated, non-pyruvylated ORF2086 polypeptide from Neisseria meningitidis serogroup B, and at least one conjugated capsular saccharide from a meningococcal serogroup, and methods of use thereof.
1. A method of producing a Neisseria meningitidis serogroup B polypeptide, comprising culturing an E. coli cell comprising a nucleic acid encoding the N. meningitidis serogroup B polypeptide set forth as SEQ. ID. NO: 55, SEQ. ID. NO: 66, SEQ. ID. NO: 68, SEQ. ID. NO: 71, SEQ. ID. NO: 76 or SEQ. ID. NO: 77 in a medium comprising glucose; disrupting the cell by microfluidization, extracting the cytoplasm of the cell; contacting the cytoplasm extract with ammonium sulfate to produce a clarified sample; loading a hydrophobic interaction column with the clarified sample; eluting the polypeptide from the hydrophobic interaction column; and purifying the eluted polypeptide by chromatography.