Virus and antigen conjugation
Various embodiments disclosed herein include methods and exemplary compositions associated with conjugation of virus and proteins (e.g., antigen) to form vaccines for delivery of immunological and other therapeutic agents, exemplary aspects of which may include harvesting viral and antigenic substances from source organisms; purifying the viral and protein (e.g. antigenic, including influenza hemagglutinin antigens) substances through methodologies described in an exemplary manner, as well as their concentration and collection; and a conjugation platform providing activation of the virus at a pH that increases binding rate and binding propensity between the virus and the protein, wherein embodiments related to the conjugation platform include controlling the ratio of virus to protein.
1. A composition comprising a virus-antigen conjugate, wherein the antigen conjugated to the virus comprises an influenza hemagglutinin antigen (HA) and exhibits at least about 50% trimer formation.
2. The composition of claim 1 , wherein the HA is chosen from the group consisting of type A HA and type B HA.
3. The composition of claim 2 , wherein the composition is multivalent.
4. The composition of claim 3 , wherein the HA comprises two or more type A hemagglutinin antigens (HAs) and two or more type B HAs.
5. The composition of claim 1 , wherein the virus is a tobacco mosaic virus (TMV).
6. The composition of claim 5 , wherein the TMV comprises surface lysine residues and the HA chemically associates with the surface lysine residues.
7. The composition of claim 6 , wherein the surface lysine residues are positioned at a N-terminal of the TMV.
8. The composition of claim 7 , wherein a carboxyl end of the HA chemically associates with the N-terminal surface lysine residues on the TMV via amine-targeted conjugation.
9. The composition of claim 5 , wherein a ratio of the TMV and the HA is expressed as TMV: HA by wt, and said ratio is in a range between 1:1 and 16:1.
10. The composition of claim 9 , wherein the ratio of the TMV and the HA is between 1:1 and 4:1.
11. The composition of claim 9 , wherein the ratio of the TMV and the HA is between 4:1 and 16:1.
12. The composition of claim 11 , wherein the ratio of the TMV and the HA is about 8:1.
13. The composition of claim 1 , wherein the virus is chosen from the group consisting of red clover mosaic virus, commelina yellow mottle virus, cauliflower mosaic virus, Soybean chlorotic mottle virus, Cassava vein mosaic virus, rice tungro bacilliform virus, petunia vein clearing virus, maize streak virus, beet curly top virus, bean golden mosaic virus, alfalfa mosaic virus, tobacco streak virus, brome mosaic virus, cucumber mosaic virus, beet yellows virus, lettuce infectious yellows virus, cowpea mosaic virus, broad bean wilt virus 1, tobacco ringspot virus, potato virus Y, ryegrass mosaic virus, barley yellow mosaic virus, parsnip yellow fleck virus, rice tungro spherical virus, carnation mottle virus, carnation ringspot virus, maize chlorotic mottle virus, tobacco necrosis virus, tomato bushy stunt virus, apple stem grooving virus, carnation latent virus, pea enation mosaic virus, soil-borne wheat mosaic virus, barley stripe mosaic virus, raspberry bushy dwarf virus, barley yellow dwarf virus, maize rayado fino virus, potato virus X, clover mosaic viruses, Southern bean mosaic virus, rice stripe virus, tobacco mosaic virus, tobacco rattle virus, apple chlorotic leaf spot virus, turnip yellow mosaic virus, and carrot mottle virus.
14. A composition comprising a virus-antigen conjugate wherein the antigen conjugated to the virus comprises an influenza hemagglutinin antigen (HA) and exhibits at least about 50% trimer formation, wherein the HA component is multivalent and comprises two or more HAs conjugated to the virus.
15. The composition of claim 14 , wherein the two or more HAs are type A HAs.
16. The composition of claim 14 , wherein the two or more HAs are type B HAs.
17. The composition of claim 14 , wherein the two or more HAs are type A or Type B HAs.