C12N2770/32151

METHOD OF PURIFYING VIRUS-LIKE PARTICLES

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.

Method of purifying virus-like-particles

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.

HANSENULA ENGINEERING FUNGI EFFICIENTLY EXPRESSING CA10 VIRUS-LIKE PARTICLES AND USE THEREOF

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.

METHOD OF PURIFYING VIRUS-LIKE-PARTICLES

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.

Recombinant foot-and-mouth disease virus with reduced immunosuppression activity, and preparation method and use thereof

The present disclosure belongs to the technical field of biological products for veterinary medicine, and specifically relates to a recombinant foot-and-mouth disease virus (FMDV) with a reduced immunosuppressive activity, a preparation method and use thereof, and a recombinant vaccine strain. According to the present disclosure, it is firstly discovered that FMDV 3B protein has an immunosuppressive function, and key sites for exerting the immunosuppressive function are found. A recombinant FMDV vaccine strain with a lost immunosuppressive function in FMDV 3B protein is constructed by introducing amino acid mutations into three repeated copies of FMDV 3B protein.

Stabilised FMDV capsids

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.

BACULOVIRUS EXPRESSION VECTOR
20230399363 · 2023-12-14 · ·

The invention concerns a baculovirus expression vector for recombinantly expressing an FMDV capsid precursor protein under control of a promoter, the expression vector comprising a nucleic acid sequence encoding the FMDV capsid precursor protein and the translational enhancers Syn2 land p10UTR. The invention further relates to a host cell comprising the baculovirus expression vector, a method of producing FMDV virus-like particles (VLPs), and a method of producing a vaccine.

Method for preparing foot-and-mouth disease vaccines
11013794 · 2021-05-25 ·

The present application relates to a method for preparing a foot-and-mouth disease (FMD) vaccine, comprising the following steps: (i) obtaining cell culture media containing FMD virus; (ii) separating and purifying the cell culture media containing FMD virus by an integrated filtration system with two membranes in combination; and (iii) collecting the concentrated solution containing FMD virus obtained in step (ii). The present application also relates to an FMD vaccine prepared by the method described herein and use thereof in the manufacture of a medicament for preventing animal FMD. The present application further relates to an apparatus for preparing an FMD vaccine, which comprises an integrated filtration system with two membranes in combination.

Processing engineered FMDV P1 polypeptide using an alternative TEV protease

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.

METHOD FOR PREPARING FOOT-AND-MOUTH DISEASE VIRUS-LIKE PARTICLES, AND TEST STRIP FOR DETECTING FOOT-AND-MOUTH DISEASE

A test strip for detecting a serotype O foot-and-mouth disease, the test strip including: a bottom board; a detection layer being disposed on the bottom board and including a detection line and a control line; an absorbent layer being disposed at one end of the detection layer close to the control line; a gold colloidal conjugate pad being disposed at the other side of the detection layer close to the detection line; and a sample pad is disposed on a top of the gold colloidal conjugate pad. The gold colloidal conjugate pad is coated with colloidal gold particles that are conjugated with a Staphylococcus protein A (SPA) marker. The detection line is coated or impregnated with serotype O FMDV-like particles, and the control line is coated or impregnated with rabbit IgG.