115 results about "Flavivirus" patented technology

Provided herein are engineered flavivirus heterotetrameric proteins (.e.g., Envelope and / or Membrane proteins) comprising at least one stabilizing amino acid substitution. The engineered flavivirus proteins form a stabilized heterotetramer structure. Messenger ribonucleic acid vaccines encoding the engineered flavivirus proteins are also provided. The disclosure also provides methods of eliciting an immune response and / or neutralizing antibodies to flavivirus in a subject.

The invention discloses an avian flavivirus NS1 trans-complementary pseudovirus system and application thereof. According to the pseudovirus system disclosed by the invention, a BHK21 cell line for stably expressing USUV-NS1 protein carrying a V5 label, namely a BHK21-NS1 stably transfected cell line, is constructed by utilizing a lentivirus packaging method; meanwhile, deletion mutation is introduced into the NS1 protein of USUV, infectious clone plasmids are constructed, and the 157th-251st amino acid of the NS1 protein is deleted by the deletion mutation. The infectious clone plasmid is transfected with a BHK21-NS1 stably transfected cell line, so that the trans-complementary virus USUV-delta NS1 which can be rescued and proliferated only in the BHK21-NS1 stably transfected cell line is obtained. The USUV-deltaNS1 trans-complementary virus is subjected to continuous passage on a BHK21-NS1 cell for 10 generations, so that the TMN-deltaNS1 virus with high yield is obtained, and complete live virus is not generated by recombination in the passage process. Therefore, the avian flavivirus NS1 trans-complementary pseudovirus system disclosed by the invention can be used as a tool to be applied to USUV vaccine preparation and related antibody evaluation.

This invention discloses the application of the DNAL1 gene as a target in screening drugs for the prevention and treatment of Zika virus or dengue virus type 2 infection, specifically the application of screening drugs for the prevention and / or treatment of Zika virus or dengue virus type 2 infection with the aim of inhibiting or knocking out the DNAL1 gene. The invention has found that the DNAL1 gene is a key host factor promoting the replication of Zika virus or dengue virus type 2, and that inhibiting the DNAL1 gene through siRNA or knocking out the DNAL1 gene through CRISPR / Cas9 can inhibit the replication of Zika virus or dengue virus type 2. The DNAL1 gene is a potential host factor for flaviviruses, and this invention provides a potential target for the prevention and / or treatment of Zika virus or dengue virus type 2 infection.

The present invention provides methods for large scale flavivirus vaccine production and manufacture. The methods provided herein are specifically contemplated for large scale production and manufacture of live attenuated flavivirus vaccines, such as live attenuated dengue virus vaccines. In addition, the methods provided herein relate to the formulation of live, attenuated, monovalent, divalent, trivalent, or tetravalent viral vaccine products.

The invention discloses establishment and application of a flavivirus replication-dependent RNA (Ribonucleic Acid) recombination experiment system, and belongs to the technical field of biology. The RNA recombination system comprises two components: a tembusu virus E gene deleted subgenome replicon CQW1-delta E, and a defective virus genome CQW1-delta NS1 or MM1775-delta NS1 with a part of NS1 gene sequence deleted. The two components cannot generate progeny viruses independently, but after the two components are co-transfected, a large number of recombinant progeny viruses can be generated within several days after transfection due to RNA recombination, the recombination efficiency is extremely high, continuous passage is not needed, the construction period is greatly shortened, and the production cost is saved; meanwhile, the generated progeny virus can form plaques on cells, and by means of plaque counting, the method can be applied to quantitative measurement of the RNA recombination efficiency and analysis of key factors influencing the tembusu virus RNA recombination.

The invention relates to the technical field of biological medicines, in particular to novel application of eltrombopag. The eltrombopag can act on an NS2B / NS3pro target spot, inhibit the activity of NS2B / NS3pro, inhibit the activity of dengue virus and Zika virus, can be applied to inhibition of the activity of flavivirus, and also has a treatment effect on diseases mediated by flavivirus.

The disclosure concerns engineered flavivirus envelope (E) polypeptides containing mutations in the stem- transmembrane domain, virus-like particles (VLPs) containing the engineered E polypeptides, and uses of the E polypeptides for treating and preventing flaviviral infections. The mutations in the stem-transmembrane domain may comprise two or more amino acid substitutions corresponding to positions 8, 11 or 15 of SEQ ID NO: 1 and the substitutions may be to a non-polar amino acid residue.

The present invention relates to polynucleotides comprising a sequence of a live, infectious, attenuated Flavivirus wherein a nucleotide sequence encoding at least a part of a Filovirus glycoprotein is located at the intergenic region between the E and NS1 gene of said Flavivirus, such that a chimeric virus is expressed, characterised in that the encoded sequence C terminally of the E protein of said Flavivirus and N terminally of the signal peptide of the NS1 protein of said Flavivirus comprises in the following order: a further signal peptide of a Flavivirus NS1 protein, a filovirus glycoprotein wherein the N terminal signal peptide is absent, a TM domain of a flaviviral E protein.

The present invention includes composition and methods for making multivalent vaccines for immunization against Flavivirus and / or arboviruses including a multivalent Virus Like Particles (VLP) and mixtures thereof, the method comprising: method of making a Flavivirus and / or arboviruses Virus Like Particles (VLP) comprising: inserting two or more nucleic acids that encode at least one Flavivirus protein into a lentiviral backbone vector; generating a lentivirus by transfecting a first cell line with the lentiviral backbone vector and isolating the lentivirus therefrom; transducing a second cell line with the lentivirus; culturing the transduced cell line under conditions in which the multivalent Flavivirus Virus Like Particles (VLP) are released from the cell line; and isolating the Flavivirus Virus Like Particles (VLP) from a culture supernatant, wherein a cell line makes a virus-specific VLP, and the VLPs are purified and then mixed in different combinations to make the multivalent vaccine.

The present invention provides compositions and methods of use comprising a stabilized recombinant E glycoprotein comprising a flavivirus E glycoprotein backbone, which comprises amino acid substitutions that stabilize the E glycoprotein in dimer conformation under physiological conditions.

The present invention relates to a method for determining the presence and / or amount of human IgG3 antibodies, preferably specific for a viral antigen, in a sample. The present invention also relates to the use of said method in the quality control of viral vaccines and in the diagnosis of viral infections. Furthermore, a kit for use in said method is described.

The application discloses a kind of establishment and application of flavivirus replication-dependent RNA recombination experimental system, belong to the field of biotechnology.The RNA recombination system includes two components: tanbusu virus E gene deletion subgenomic replicon CQW1-ΔE, and the defective virus genome CQW1-ΔNS1 or MM1775-ΔNS1 missing partial NS1 gene sequence.The two components cannot produce progeny virus alone, but after co-transfection, due to RNA recombination, a large number of recombinant progeny viruses can be produced within a few days after transfection, the recombination efficiency is extremely high, and continuous passage is not required, greatly shortening the construction cycle and saving production costs;At the same time, the progeny virus can form plaques on cells, and the efficiency of RNA recombination can be quantitatively measured by plaque counting, and the key factors affecting tanbusu virus RNA recombination can be analyzed.

The present invention relates to an isolated mosquito extracellular vesicle for use as an immunogenic agent in the prevention of at least one vector-borne disease. The present invention also relates to a vaccine composition comprising at least one mosquito extracellular vesicle. The present invention further relates to the use of a mosquito extracellular vesicle in identifying at least one immunogenic peptide capable of reducing, in a vertebrate, the transmission of a vector-borne disease when the vertebrate is bitten by the vector, and to a method for identifying at least one immunogenic peptide capable of blocking, in a vertebrate, the transmission of a vector-borne disease when the vertebrate is bitten by the vector, the method comprising a step of identifying in vitro at least one target antibody present in an animal serum sample immunised against mosquito extracellular vesicles.

The invention provides a discovery method and application of a flavivirus vaccine antigen. According to the flavivirus vaccine antigen discovery method disclosed by the invention, the flavivirus vaccine antigen with damaged FL epitope and normal prME protein expression and folding can be discovered with relatively high efficiency, and the obtained flavivirus vaccine antigen can avoid an ADE effect, has relatively good immunogenicity and can be used for constructing flavivirus vaccines; therefore, the discovery method of the flavivirus vaccine antigen can be used for development of flavivirus vaccines, and has an excellent industrialization prospect.

Ligands that target Flavivirus; conjugates comprising radicals of such ligands; compositions comprising the conjugates; methods of use of the ligands and / or conjugates to target active agents to the Flavivirus E protein; and methods of use of the conjugates to treat and prevent Flavivirus infection.

Compounds, compositions and methods for preventing, treating or curing a coronavirus infection in human subjects or other animal hosts. In one embodiment, the compounds can be used to treat an infection with a severe acute respiratory syndrome virus, such as human coronavirus 229E, SARS, MERS, SARS-CoV-1, OC43, and SARS-CoV-2. In another embodiment, the methods are used to treat a patient infected with a Flavivirus, Picornavus, Togavirus, or Bunyavirus.

The invention relates to the technical field of biomedicine, and provides a new target, a new small nucleic acid drug and application of the new small nucleic acid drug in preparation of drugs for prevention or treatment for resisting infection of three viruses of Zika virus (ZIKV), dengue virus (DENV) and hepatitis C virus (HCV). According to the invention, the specific nucleotide sequences of the target lipid droplet coating protein (Perilipin), which are shown as SEQ ID NO.1 to SEQ ID NO.3, are obtained through screening. Experiments discover that Perilipin plays an important role in flavivirus infected target cells, expression and replication of the three virus proteins and genome RNA in the target cells can be effectively blocked by down-regulating expression of Perilipin, and therefore it is proved that Perilipin can serve as a new target spot for resisting infection of the three flaviviruses, and the application of Perilipin in resisting flavivirus infection is proven to be a new target spot for resisting infection of the three flaviviruses. Specific small nucleic acid can be used as an antiviral drug for the three flaviviruses.

The present invention provides methods for large scale production and manufacturing of flavivirus vaccines. The methods provided herein are particularly contemplated for large scale production and manufacturing of live attenuated flavivirus vaccines, such as live attenuated dengue virus vaccines. Additionally, the methods provided herein relate to the formulation of live attenuated, monovalent, bivalent, trivalent, or quadrivalent viral vaccine products.

The invention belongs to the technical field of microbial viruses, and particularly relates to a culture method for improving the virus titer of duck flavivirus, which comprises the following steps: step 1, preparing a culture solution, accurately weighing RPMI1640 dry powder, dissolving the RPMI1640 dry powder in three-distilled water, then adding a proper amount of fetal calf serum and double antibodies, filtering and sterilizing, then sub-packaging into sterile culture bottles, and preparing the culture solution for later use; storing at a proper temperature for later use; 2, preparing cells, selecting suitable cells, inoculating the cells into a culture flask, and carrying out single-layer culture by using an RPMI1640 culture solution to ensure that the cells are in a good growth state before the cells are inoculated with viruses, The method has the beneficial effects that by selecting a suitable host cell line, the cells are ensured to be in a healthy state, the performance reduction caused by high algebraic culture is avoided, and the cell quality is improved. In addition, the cell culture medium suitable for virus production is selected, and the culture medium with high transfection efficiency and cell maintaining capacity after transfection can remarkably improve the virus titer.

This disclosure relates to stabilizers for compositions comprising live-attenuated flaviviruses, methods for stabilizing live-attenuated flaviviruses, and stabilized live-attenuated flavivirus vaccine compositions.

Compounds, methods, and compositions for treating or preventing viral infections using nucleoside compounds. The nucleoside compounds have increased antiviral activity potential with the result of inhibiting at least one of flaviviruses, herpesviruses, polyomaviruses, alphaviruses, enteroviruses, filoviruses matonaviruses, phenuiviruses, Hepatitis B virus, and / or coronaviruses.

The application belongs to the field of biological pharmacy, and provides application of a compound or a derivative or a salt thereof in prevention and treatment of vertical transmission of viruses, in particular, provides use of a compound or a pharmaceutically acceptable salt thereof in preparation of a medicine, and the medicine is used for at least one of the following: (1) inhibition and / or blocking of transmission of flavivirus from a mother to an embryo and / or a fetus; (2) prevention and / or treatment of flavivirus infection of the embryo and / or the fetus, and diseases caused by the flavivirus infection, wherein the compound comprises a transcytosis pathway inhibitor. The compound or the pharmaceutically acceptable salt thereof cannot significantly reduce the load of the flavivirus in the mother, but can effectively and specifically reduce the amount of the flavivirus infection in the fetus and the pre-embryo, significantly inhibits the vertical transmission of the flavivirus in the fetus or the embryo after the flavivirus infects the mother, and effectively guarantees the safety during pregnancy.

The present invention pertains to compounds that inhibit Flavivirus NS2B-NS3 proteases and are potent and selective inhibitors of the West Nile virus and Zika virus. The compounds of the invention can be used alone or in combination with other agents to treat infections caused by these viruses. Additionally, the invention relates to pharmaceutical compositions containing these compounds.

The present disclosure provides nanostructures and nanostructure-based vaccines. Some nanostructures of the present disclosure display antigens capable of eliciting an immune response to infectious agents such as bacteria, viruses, and pathogens. Some vaccines of the present disclosure are useful for preventing or reducing the severity of infection by infectious agents including, but not limited to, Lyme disease, pertussis, herpes virus, orthomyxovirus, paramyxovirus, pneumovirus, filovirus, flavivirus, reovirus, retrovirus, meningococcus, or malaria. The antigens can be attached to the core of the nanostructure non-covalently or covalently, including as a fusion protein or by other means disclosed herein. Multimeric antigens can optionally be displayed along the symmetry axis of the nanostructure. Also provided are proteins and nucleic acid molecules encoding such proteins, vaccine compositions, and methods of administration.

The invention provides a vaccine composition comprising a flavi peptide comprising one or more CD8+ T cell epitopes.

The invention relates to a technical method for quickly and forcibly moulting cherry valley meat breed ducks. The method comprises the following steps: on the first day, cutting off water and feed for a whole group, picking and panning, grouping, fixing a fence and weighing; on the second day, the male ducks are not supplied with water, and the female ducks are supplied with water for 1 h and not supplied with water; on the third day, supplying 120g of feed to each male duck in the egg producing period, supplying water to each female duck for 1 hour in the morning and afternoon, and not supplying the feed; from the fourth day to the sixth day, the disabled ducks are selected and washed, and antibiotics are continuously fed for 3 days; from the seventh day to the tenth day, weightlessness is calculated through weighing, and when the weightlessness is 30%, main wing feathers and tail feathers are pulled out; from the 11th day to the 28th day, 50 g of feed is supplied to each female duck in the egg producing period, then 5 g to 140 g of feed is supplied to each female duck every day, illumination is conducted for 13 h, and then 1 h to 17 h is increased every week; on the 29th day, weighing 10% of each fence, grouping in 30 days when the weight reaches the standard, and grouping in 37 days when the weight is lower than the standard; on the 30th day or the 37th day, the residual ducks are selected and washed, the ducks are combined, and antibiotic fallopian tubes are fed for purification; on the 25th day, the 32nd day, the 39th day and the 46th day, flavivirus, H9, H5 + H7, duck liver + duck plague are prevented.

The present invention relates to the field of biopharmaceutical technology, and more specifically, to a novel use of eltrombopag. Eltrombopag can act on the NS2B / NS3pro target site, inhibiting the activity of NS2B / NS3pro, while also inhibiting the activity of dengue and Zika viruses. It can also be used to inhibit the activity of flaviviruses and has therapeutic effects on diseases mediated by them.

Ligands that target Flavivirus; conjugates comprising radicals of such ligands; compositions comprising the conjugates; methods of use of the ligands and / or conjugates to target active agents to the Flavivirus E protein; and methods of use of the conjugates to treat and prevent Flavivirus infection.

The invention relates to the technical field of plant virus detection, in particular to a pepper vein flavivirus detection method and system based on RT-RAA-CRISPR / Cas12a and application of the pepper vein flavivirus detection method and system. The invention discloses a pepper vein flavivirus detection method based on RT-RAA-CRISPR / Cas12a. The pepper vein flavivirus detection method comprises the following steps: S1, extracting virus RNA (Ribonucleic Acid) from a plant leaf to be detected; s2, adding RNA (Ribonucleic Acid) of a sample to be detected into the RT-RAA amplification system, and carrying out isothermal amplification at 37-42 DEG C for 10-30 minutes to obtain an RT-RAA amplification product; s3, adding the RT-RAA amplification product into a CRISPR / Cas12a (clustered regularly interspaced short palindromic repeats / CRISPR associated 12a) detection system, and reacting for 20 to 30 minutes under the condition of 37 DEG C; s4, detecting fluorescence change in real time, and judging a result through a fluorescence signal; if a fluorescence signal is detected, it is indicated that the pepper vein flavivirus exists in the RNA of the to-be-detected sample, efficient amplification is achieved through a recombinase-single-chain binding protein-primer compound, and high sensitivity and specificity of detection are ensured through accurate nucleic acid recognition and signal amplification functions; the method is especially suitable for rapid screening and diagnosis of plant viruses in limited resource environments such as fields.

The invention relates to the field of enzyme engineering and molecular biology, and discloses a heterologous virus-derived chimeric multifunctional 5 '-'capping' enzyme, and the mRNA vaccine 5 '-'capping' enzyme is formed by sequentially connecting an RNA triphosphatase functional domain derived from rabbit fibroma virus, a guanylyl transferase functional domain and a methyltransferase functional domain derived from flavivirus NS5 protein. According to the invention, RNA triphosphatase, guanylyl transferase and a methyltransferase structural domain with a dual methylation function are integrated into a single polypeptide chain, so that a catalytic process of completing synthesis of a 5'cap 'structure of the mRNA vaccine by one enzyme and one step is realized, the modification process flow of the mRNA vaccine is remarkably simplified, and the consistency and reproducibility of mRNA vaccine production are improved.