3.2 Stability, functionality, and immune response is preserved after storage for 6 months
Plant seeds has natural ability to protect the vaccine antigen by bioencapsulation in the plant tissues, and therefore allow its storage at ambient temperature, moisture, and light conditions, and thus simplifies its production and distribution. We have tested our plant made vaccine antigen for confirmation of immunogenicity after long-term storage. The immunoblot (Fig. 3) confirmed TM-1 vaccine antigen can be stored at ambient temperature without degradation for at least six months. The confirmation of immune response is evident of such bioencapsulation where vaccine antigen was intact during passing through digestive system. Cell wall mediated bioencapsulation allow proteins to withstand enzymatic condition and can reach the small intestine, where the TM-1 vaccine antigen was incorporated into M cells in the follicle-associated epithelium (FAE), and finally to systemic immune responses. The CTB is FDA approved as fusion protein for delivery of biopharmaceuticals into immune system. The furin sites are incorporated between CTB and TM1 antigen for its delivery upon its cleavage. In addition, such results are encouraging because this protection to withstand biodegradation elements and stability at normal room conditions would allow plant made vaccine to eliminate the moderate to ultra-cold conditions required for vaccine storage and distribution. Also, we confirmed the feasibility of oral delivery of plant made vaccine production and its efficacy for animals with efficient delivery vehicle such as CTB and cleavage sites