The present disclosure relates to hand, foot, and mouth disease vaccines and immunogenic compositions having one or more antigens from at least one virus that causes hand, foot, and mouth disease in humans, and methods of manufacture, formulation, and testing, and uses thereof.

A method of producing purified FMDV VLPs, comprising contacting cells containing FMDV VLPs with a lysis buffer and allowing the cells to lyse, the lysis buffer comprising 10-20 mM Tris-HCl, 150-200 mM NaCl, 3 mM MgCl.sub.2, and 1% Triton X-100, wherein the lysis buffer does not contain EDTA; centrifuging a solution; and removing a supernatant from the solution, the supernatant containing the purified FMDV VLPs.

A method of producing purified FMDV VLPs, comprising contacting cells containing FMDV VLPs with a lysis buffer and allowing the cells to lyse, the lysis buffer comprising 10-20 mM Tris-HCl, 150-200 mM NaCl, 3 mM MgCl.sub.2, and 1% Triton X-100, wherein the lysis buffer does not contain EDTA; centrifuging a solution; and removing a supernatant from the solution, the supernatant containing the purified FMDV VLPs.

The present invention provides EV71 virus-like particles and a preparation method and application thereof. The method comprises: connecting a P1 protein gene and a 3CD protease gene of an EV71 virus with a PMV plasmid to construct a PMV-P1-3CD recombinant expression plasmid; then transforming a Hansenula polymorpha AU-0501 expression strain with the PMV-P1-3CD recombinant expression plasmid to obtain an AU-PMV-P1-3CD recombinant expression strain; fermenting and culturing the recombinant expression strain, and inducing the recombinant expression strain to express the EV71 virus-like particle protein with methanol; centrifuging and collecting mycelia for homogeneous breakage at a high pressure; and purifying the supernatant through ion-exchange chromatography, hydrophobic chromatography, and molecular sieve chromatography, so as to obtain EV71 virus-like particles.

The present invention provides EV71 virus-like particles and a preparation method and application thereof. The method comprises: connecting a P1 protein gene and a 3CD protease gene of an EV71 virus with a PMV plasmid to construct a PMV-P1-3CD recombinant expression plasmid; then transforming a Hansenula polymorpha AU-0501 expression strain with the PMV-P1-3CD recombinant expression plasmid to obtain an AU-PMV-P1-3CD recombinant expression strain; fermenting and culturing the recombinant expression strain, and inducing the recombinant expression strain to express the EV71 virus-like particle protein with methanol; centrifuging and collecting mycelia for homogeneous breakage at a high pressure; and purifying the supernatant through ion-exchange chromatography, hydrophobic chromatography, and molecular sieve chromatography, so as to obtain EV71 virus-like particles.

The present invention provides for a novel oil-in-water (O/W) emulsion, with increased stability in the presence of bacterial or viral suspensions, especially those concentrated and non-purified (crude extracts) or minimally purified. The emulsion of the present invention can act as vehicle for the delivery of a pharmaceutical composition comprising at least one immunogen and, in particular, an immunogen selected from the group consisting of an inactivated pathogen, an attenuated pathogen, a subunit, a recombinant expression vector, and a plasmid or combinations thereof.

The present invention provides for a novel oil-in-water (O/W) emulsion, with increased stability in the presence of bacterial or viral suspensions, especially those concentrated and non-purified (crude extracts) or minimally purified. The emulsion of the present invention can act as vehicle for the delivery of a pharmaceutical composition comprising at least one immunogen and, in particular, an immunogen selected from the group consisting of an inactivated pathogen, an attenuated pathogen, a subunit, a recombinant expression vector, and a plasmid or combinations thereof.

Provided herein are adenoviral nucleic acid sequences and adenoviral vectors comprising said nucleic acid sequences. The provided adenoviral vectors can be used to induce a protective immune response in a subject.

Provided herein are adenoviral nucleic acid sequences and adenoviral vectors comprising said nucleic acid sequences. The provided adenoviral vectors can be used to induce a protective immune response in a subject.

The present invention provides attenuated P. multocida strains that elicit an immune response in animal P. multocida, compositions comprising said strains, methods of vaccination against P. multocida, and kits for use with such methods and compositions. The invention further provides novel, genetically-engineered mutations in P. multocida hyaD and nanPU genes, which are useful in the production of novel attenuated P. multocida bacterial strains.