Haemophilus influenzae vaccine and methods of use

A method of making a fusion protein comprising all or part of two or more Haemophilus influenzae (Hi) proteins selected from the group consisting of Omp26, P6, PD and PF, wherein at least one of the Hi proteins is lipidated, the method comprising:

• i) inserting a nucleic acid encoding a lipid moiety signal sequence region upstream of a first nucleic acid encoding all or part of a Haemophilus influenzae (Hi) protein Omp26, P6, PD or PF in a plasmid vector;
• ii) inserting a second nucleic acid encoding all or part of a Haemophilus influenzae (Hi) protein Omp26, P6, PD or PF in the plasmid vector;
• iii) optionally inserting a third or additional nucleic acid sequences encoding one or more additional Hi proteins or fragment thereof selected from the group consisting of Omp26, P6, PD and PF;
• iii) transfecting the plasmid vector into a host cell capable of expressing the nucleic acid molecules;
• and
• iv) purifying the fusion protein expressed by the plasmid.

A method of making a fusion protein comprising all or part of two or more Haemophilus influenzae (Hi) proteins selected from the group consisting of Omp26, P6, PD and PF, wherein at least one of the Hi proteins is lipidated, the method comprising:

• i) providing a nucleic acid encoding a lipid moiety signal sequence region;
• ii) providing a first nucleic acid encoding all or part of a Hi protein Omp26, P6, PD or PF;
• iii) providing a second nucleic acid encoding all or part of a Hi protein Omp26, P6, PD or PF, wherein the second nucleic acid encodes a different Hi protein from i);
• iv) optionally providing a third or additional nucleic acid sequences encoding one or more additional proteins or fragment thereof selected from the group consisting of Omp26, P6, PD and PF;
• v) inserting the nucleic acid sequences i-iv) into a plasmid vector capable of expressing the nucleic acids;
• vi) transfecting the plasmid vector of iv) into a host cell capable of expressing the nucleic acid molecules and expressing the fusion protein;
• and
• vii) purifying the recombinant fusion protein expressed in the host cell.