Composition for oral or nasal delivery of tetanus, diphtheria, and pertussis vaccine alone or in combination using neurotoxin associated proteins

A process for the preparation of a vaccine and neurotoxin associated proteins (NAPs) composition, comprising:

• mixing an equimolar ratio of the vaccine and neurotoxin associated proteins (NAPs) into a mixture that is maintained at a pH of from 5.5 to 6.2,
• wherein the vaccine comprises a detoxified recombinant protein(s) derived from one or more proteins associated with an infectious disease selected from the group consisting of tetanus, diphtheria, pertussis alone or in combination thereof;
• wherein the neurotoxin associated proteins (NAPs) are derived from Clostridium botulinum or botulinum neurotoxin, and the NAPs become chemically or recombinantly linked or otherwise associated to the vaccine during the mixing;
• maintaining the mixture at the pH in the range of from 5.5 to 6.2 using 25 mM phosphate buffer at a pH 5.8 containing 200 mM sodium chloride solution;
• reacting the mixture with at least 10 mM imidazole solution at an ambient temperature for from 2 to 3 hours;
• loading the mixture to a Nickel-Nitrilotriacetic acid (Ni-NTA) affinity column;
• and
• eluting the column to get a pure vaccine and neurotoxin associated proteins (NAPs) composition.