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 second-generation tetanus toxoid vaccine, comprising the steps of:

• a) inducing an E. coli culture at OD600=0.5 by adding 0.2 mM isopropyl-β-D-1-thiogalactopyranoside (IPTG);
• b) growing the culture at 14-16° Celsius (C) for 14 to 20 hours;
• c) suspending the culture in 25 mM phosphate buffer containing 200 mM sodium chloride;
• d) adding 1% of (2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol) to the phosphate buffer and resuspending the culture in said phosphate buffer;
• e) sonicating the culture for a period of 3 minutes (at 5 sec on/off pulse) at 4° C. on cold beads;
• f) centrifuging the culture for 45 to 90 minutes;
• g) collecting and purifying the supernatant using Nickel-Nitrilotriacetic acid (Ni-NTA) affinity column with an eluant;
• and
• h) combining the eluant into a pool with contaminated bands and concentrating using centrifugal filters to isolate one or more detoxified domains of a tetanus neurotoxin (DrTeNT) heavy chains proteins.