The present disclosure provides a type A foot-and-mouth disease virus-like particle antigen assembled by VP2, VP3 and VP1 antigen proteins of an epidemic strain of type A foot-and-mouth disease virus. The type A foot-and-mouth disease virus VP2 antigen protein is encoded by a nucleotide sequence shown in SEQ ID No. 1 or its degenerate sequence, the type A foot-and-mouth disease virus VP3 antigen protein is encoded by a nucleotide sequence shown in SEQ ID No. 2 or its degenerate sequence, and the type A foot-and-mouth disease virus VP1 antigen protein is encoded by a nucleotide sequence shown in SEQ ID No. 3 or its degenerate sequence.

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 a Hansenula engineering fungus that efficiently expresses CA10 virus-like particles and use thereof. The engineering fungus comprises a recombinant vector carrying P1 and 3CD genes of the CA10 virus, and the starting strain of the engineering fungus is uracil auxotroph. The Hansenula AU-0501, P1 and 3CD genes are optimized according to preferred codons of Hansenula. The present invention also provides a preparation method of CA10 virus-like particles, comprising: culturing engineering fungi, expressing CA10 virus-like particles, and separating and purifying virus-like particles by ultrafiltration and three-step chromatography. The CA10 virus-like particles provided by the present invention and the vaccine prepared therefrom have good immunogenicity, safety and biological activity, the process is simple, the chromatography method is adopted for purification, and large-scale preparation and purification can be realized to obtain a VLP protein stock solution with high purity (greater than 99%), which can be used to prepare a vaccine to prevent CA10 infection, with good economic value and application prospects.

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.

Provided is a method to generate a recombinant virus that can stably express target proteins. The recombinant virus of the present invention is useful for producing an immunogenic composition or vaccine.

The present disclosure provides a type O foot-and-mouth disease virus-like particle antigen, wherein the type O foot-and-mouth disease virus-like particle antigen is type O CATHAY type foot-and-mouth disease virus-like particle antigen, and the type O CATHAY type foot-and-mouth disease virus-like particle antigen is assembled by VP0, VP3 and VP1 antigen proteins of type O CATHAY type foot-and-mouth disease virus. The type O foot-and-mouth disease virus-like particle antigen of the present disclosure has good immunogenicity. The prepared vaccine can produce complete protection against the O-type foot-and-mouth disease virus on the 14th day after immunization. The antibody titer produced is higher than that of the commercial inactivated vaccine, and the duration of immune protection can be maintained for at least 133 days. The disclosure also relates to the prepared vaccine composition, preparation method and use thereof.

The present invention relates to the stabilisation of foot-and-mouth disease virus (FMDV) capsids, by specific substitution of amino acids in a specific region of FMDV VP2. The invention provides stabilised FMDV capsids and vaccines against FMD.

Polynucleotide constructs that express an engineered foot-and-mouth disease (FMDV) P1 precursor protein and a non-FMDV TEV protease and methods for safe and efficient recombinant production of FMDV antigens and immunogens. Recombinant production of FMDV antigens avoids the need to culture highly-infectious FMDV, while conventional culture methods for producing FMDV antigens rely on the native FMDV 3C protease which exerts toxic effects on host cells. The inventors have developed a new system that efficiently and safely processes FMDV P1 precursor without the FMDV 3C protease, thus avoiding the toxic effects associated with use of the 3C protease. The invention is also directed to the FMDV antigens and virus-like particles produced by this system as well as to FMDV vaccines, diagnostics and other biologics.

This application is directed generally to foot-and-mouth disease virus (FMDV) 3C proteases that have been modified by mutating a polynucleotide sequence coding for the FMDV 3C protease. The modified FMDV proteases exhibit proteolytic activity on FMDV P1 precursor protein and exhibit a reduction in one or more toxic or inhibitory properties associated with an unmodified FMDV 3C protease on a host cell used to recombinantly produce it. Vectors carrying polynucleotides encoding modified FMDV 3C protease sequences can induce production of FMDV virus-like particles in a host cell when expressed in the host cell. The modified FMDV 3C proteases can generally be used to produce immunogenic FMDV preparations capable of inducing an immune response against FMDV.