173 results about "Rabies virus" patented technology

The invention discloses a pseudorabies virus variant gD protein as well as a preparation method and application thereof. The amino acid sequence of the pseudorabies virus variant gD protein is shown as a sequence 1 in a sequence table, and the nucleotide sequence of the pseudorabies virus variant gD protein is shown as a sequence 2 in the sequence table. The pseudorabies virus variant gD protein is used for indirect ELISA, can be used for detecting whether a pseudorabies virus variant gD protein antibody exists in pig blood or not, and is simple, convenient and rapid in detection, high in specificity, high in sensitivity, accurate and reliable. The pseudorabies virus variant gD protein can be prepared into a subunit vaccine, can be induced to generate a high-level anti-pseudorabies virus variant neutralizing antibody after immunizing a pig, has the advantages of strong immunogenicity, high safety and matching with a current epidemic strain, can distinguish vaccine immunization and wild virus infection animals through differential diagnosis, and has a wide application prospect. The method can be used for preventing and controlling the propagation and prevalence of pseudorabies. The gD protein of the pseudorabies virus variant can be used for detecting and diagnosing pseudorabies and preventing pseudorabies, and has good production and application prospects in development of novel diagnostic reagents and subunit vaccines of pseudorabies viruses.

The invention relates to the technical field of biological medicines, in particular to a recombinant rabies virus nano-antibody fused with a solubilizing oligopeptide and a cell penetrating peptide and application of the recombinant rabies virus nano-antibody in preparation of rabies prevention and treatment products. The recombinant rabies virus nano-antibody fused with the solubilizing oligopeptide and the cell penetrating peptide comprises a humanized rabies virus nano-antibody, and the C end of the humanized rabies virus nano-antibody is fused with a solubilizing tag peptide P17 and the cell penetrating peptide. The recombinant rabies virus nano antibody utilizes a prokaryotic expression system, is low in cost, has excellent cell membrane and BBB penetrating ability, can enter a central nervous system from veins for virus neutralization treatment, and has an efficient passive immune effect.

The present invention provides a rabies virus G protein antigen epitope targeting completely human monoclonal antibody and applications thereof, the completely human monoclonal antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region has three complementary determining region amino acid sequences: GDISSSCFY, IHYSGST and ARHRRGYCYDSEKGGTNWFDP, and the light chain variable region has three complementary determining region amino acid sequences: GDISSSCFY, IHYSGST and ARHRRGYCYDSEKGGTNWFDP. The amino acid sequences of the three complementary determining regions of the light chain variable region are respectively as follows: QGISND, ATS and LQDYEFPLT. The fully human monoclonal antibody has efficient and broad-spectrum anti-rabies virus neutralizing activity, is high in expression, fully human-derived and good in stability, and can be used for preparing rabies virus detection products or drugs for preventing and treating rabies.

Polypeptides comprising an extracellular domain of rabies virus G protein are provided. The rabies virus G protein extracellular domain may have a deletion of the fusion ring domain. Further provided are nanoparticle vaccines for rabies virus. Further provided are pharmaceutical compositions, methods of making, and methods of using, such as immunizing a subject to produce a protective immune response against rabies virus.

The invention relates to a novel rabies virus nucleocapsid nanostructure vaccine and a construction method thereof, and belongs to the field of biomedicine.The nanostructure vaccine is characterized in that mRNA capable of efficiently expressing rabies virus G protein and recombinant rabies virus N protein expressed by escherichia coli are combined to form an NLS structure serving as a vaccine antigen component; and then wrapping with a lipid component. The vaccine constructed by the invention can form G antigen and N antigen integrated immunogen signal stimulation, so that specific immunity can form an effective reaction in humoral immunity and cellular immunity. In a protective test of challenge, the vaccine can achieve a 100% protection effect on mice.

The invention discloses a recombinant rabies virus genome plasmid for high-sensitivity expression of red fluorescent protein, a recombinant rabies virus as well as a preparation method and application of the recombinant rabies virus. The preparation method specifically comprises the following steps: (1) constructing the recombinant rabies virus genome plasmid for high-sensitivity expression of red fluorescent protein as shown in SEQ ID NO.2; (2) preparing a recombinant rabies virus for expressing the red fluorescent protein with high sensitivity; and (3) application of the recombinant rabies virus for high-sensitivity expression of the red fluorescent protein in neural circuit tracing. The recombinant rabies virus capable of highly sensitively expressing the red fluorescent protein can efficiently mark an upstream neural network in a retrograde manner, so that the problem of low efficiency of expressing red fluorescence in the existing method is solved; the method has wide application value and market prospect in the aspects of neural circuit structure and function analysis, neuron sparse marking, vaccine and drug research and development, disease model establishment, neural network loss and reconstruction detection, virus replication or pathogenic mechanism research and the like.

The invention provides a rabies virus inactivated vaccine and a preparation method thereof, and belongs to the technical field of biological medicines. The invention provides a rabies virus inactivated vaccine. The rabies virus inactivated vaccine comprises an adjuvant and an inactivated VSV-rabies recombinant virus, the inactivated VSV-rabies recombinant virus is obtained by carrying out inactivation on a VSV-rabies recombinant live virus; the VSV-rabies recombinant live virus is prepared by replacing envelope protein G of vesicular stomatitis virus with G protein of rabies virus. The inactivated rabies virus vaccine is good in safety and high in induced antibody level, the immune effect of the inactivated rabies virus vaccine is equivalent to that of a live virus vector vaccine, and the inactivated rabies virus vaccine has a certain application value in the aspect of preventing and controlling rabies.

The present invention discloses a combined monoclonal antibody preparation against rabies virus, which relates to the technical field of biomedicine. The combined monoclonal antibody preparation against rabies virus contains R92 antibody and R71 antibody; the light chain sequence of the R92 antibody is as shown in SEQ ID NO:1, the heavy chain sequence of the R92 antibody is as shown in SEQ ID NO:2, the light chain sequence of the R71 antibody is as shown in SEQ ID NO:3, and the heavy chain sequence of the R71 antibody is as shown in SEQ ID NO:4. It is also disclosed that the ratio of the R92 antibody to the R71 antibody in the combined monoclonal antibody preparation against rabies virus is (1-5):(1-5). The combined monoclonal antibody preparation against rabies virus of the present invention has a high affinity for rabies virus glycoprotein, has good neutralization ability and thermal stability, has good neutralization ability against a variety of street strains, and the two antibodies target different epitopes, providing a wider range of protection.

The invention discloses preparation and application of a rabies virus G protein mRNA vaccine freeze-drying preparation, and relates to the field of veterinary biological products. According to the invention, rabies virus G protein mRNA is prepared on a large scale through an in-vitro transcription technology, and an mRNA-LNP (lipid nanoparticle) delivery system is successfully constructed by adopting a microfluidic encapsulation process. Through freeze-drying protective agent formula screening (including cane sugar, trehalose and mannitol) and freeze-drying procedure parameter optimization (including pre-freezing temperature, main drying rate and secondary drying residual moisture control), the freeze-dried mRNA vaccine preparation which can stably exist under the condition of 4 DEG C is finally obtained.

An anti-rabies virus neutralizing antibody and preparation and a use thereof. Provided is an anti-rabies virus neutralizing antibody, comprising a heavy chain variable region SEQ ID NO. 1-SEQ ID NO. 3, a light chain variable region SEQ ID NO. 4-SEQ ID NO. 5, and GAS. The anti-rabies virus neutralizing antibody has good neutralizing activity; a high isoelectric point provides advantages in the stability of antibody formulations; a high Tm value indicates good thermal stability; strong affinity with recombinant rabies virus glycoproteins shows promise for the mass production of rabies virus neutralizing antibodies, providing technical support for the preparation of low-cost, safe, and effective rabies passive immune formulations.

The invention relates to circRNA (Ribonucleic Acid) for coding rabies virus as well as a vaccine and application thereof, and belongs to the technical field of biological medicines. The invention relates to circRNA (Ribonucleic Acid) for coding rabies virus. The nucleotide sequence of the circRNA is shown as SEQ ID NO: 1. The circRNA comprises an SRV9 strain rabies virus G protein sequence after coding optimization, the circRNA is delivered through lipid nanoparticles to form the circRNA-LNP vaccine, powerful and lasting body fluid and cellular immune response can be induced in mice and non-human primates, and the complete protection effect can be achieved through a single dose. Compared with a traditional inactivated vaccine, the vaccine has the advantages of being rapid in production, high in heat stability, capable of inducing Th1 type immune bias and the like, is suitable for preventing rabies virus infection and has wide application prospects and popularization value.

The invention provides a perfusion culture preparation method and application of a recombinant human anti-rabies virus monoclonal antibody R71, and relates to the technical field of biological medicines. According to the method, key process parameters in the culture process of the engineering cell strain expressing the R71 antibody are optimized, the key processes comprise a culture medium, a rotating speed, pH, an ATF P-flow set value and a dissolved oxygen (DO) range, and experimental results show that the receiving amount of R71 in each batch of the optimized perfusion culture method can reach more than 200g and is 5-8 times that of fed-batch culture under the same culture scale.

The invention discloses an anti-rabies virus N protein monoclonal antibody, a detection reagent and application of the anti-rabies virus N protein monoclonal antibody. The monoclonal antibody for resisting the rabies virus N protein comprises a heavy chain variable region and a light chain variable region, the amino acid sequence of the heavy chain variable region is as shown in SEQ ID No. 1; the amino acid sequence of the light chain variable region is as shown in SEQ ID No.2. The monoclonal antibody with relatively high affinity and detection sensitivity for resisting rabies virus N protein lays a foundation for research, development and popularization of colloidal gold test strips and fluorescence immunoassay test strips.

The invention discloses a construction method and application of a rabies virus RNA polymerase dual-luciferase report system, and belongs to the technical field of biology. In order to solve the technical problem that a report system for detecting the RABV RNA polymerase activity needs to be developed urgently, a Firefly luciferase fragment and a vector pcDNA3.1-SRV9-M-G obtained by amplifying M protein and G protein-deleted rabies virus SRV9 strain full-length plasmid are seamlessly connected, a recombinant luciferase report plasmid is constructed, the recombinant luciferase report plasmid and an auxiliary plasmid are combined, and the recombinant luciferase RNA polymerase activity detection system is constructed. And the RABV RNA polymerase dual-luciferase report system is formed by the Renilla luciferase report plasmid and the T7 promoter plasmid, and the RABV RNA polymerase dual-luciferase report system comprises the Renilla luciferase report plasmid and the T7 promoter plasmid. The system can accurately and efficiently detect the activity of RABV polymerase, and by virtue of the function, effective antiviral targets and therapeutic drugs can be screened out in an assisted manner.

The present disclosure provides a recombinant vector from an attenuated rabies virus comprising a nucleotide sequence encoding at least one CCHFV glycoprotein and at least one mucin like domain. The disclosure further provides a method for conditioning an immune response with said recombinant virus vector incorporated in said virus virion.

The invention discloses application of siRNA for targeted inhibition of rabies virus G gene and / or L gene in preparation of drugs for inhibiting rabies. SiRNA-G for targeted inhibition of a rabies virus G gene and siRNA-L for targeted inhibition of a rabies virus L gene are provided on the basis, the nucleotide sequence of a sense strand of the siRNA-G is as shown in SEQ ID NO: 1, and the nucleotide sequence of an antisense strand of the siRNA-G is as shown in SEQ ID NO: 2; the nucleotide sequence of the positive-sense strand of the siRNA-L is as shown in SEQ ID NO: 3, and the nucleotide sequence of the antisense strand of the siRNA-L is as shown in SEQ ID NO: 4. The siRNA-G and / or siRNA-L can effectively inhibit the expression of the G gene and the L gene of the rabies virus and reduce the proliferation of the rabies virus, and can be used as an effective small nucleic acid drug in the development process of anti-rabies virus drugs.

The invention belongs to the technical field of vaccine preparation, and particularly relates to a method for suspension culture of rabies virus. MDCK-siat7e-C is used as a cell matrix for suspension culture of the rabies virus, experiments prove that the rabies virus can be subcultured in the cell, the virus titer can reach 7-81gFFU / ml, and through mouse immunogenicity inspection, the protection index is greater than 100, and the antibody titer reaches 67IU / ml after mice are immunized. The method for suspension culture of the rabies virus by the cell strain provides reference for suspension culture of the rabies virus.

The invention discloses application of mitoxantrone in preparation of an anti-rabies virus drug, reveals that clinical anti-tumor and immunosuppression drug mitoxantrone has remarkable anti-rabies virus activity for the first time, and opens up a new application field for mitoxantrone. The rigorous in-vitro and in-vivo tests prove that the mitoxantrone can inhibit virus replication in a dose-dependent manner, reduce virus gene and protein expression and remarkably improve the survival rate of infected mice in an animal model. The invention provides a new clinical application approach for mitoxantrone, provides a powerful candidate drug and a brand-new technical scheme for anti-rabies virus drugs, and has important practical significance and clinical application prospects for coping with rabies which is a major public health threat.

The present invention relates to the field of biomedicine, and more particularly to a method for purifying an inactivated rabies virus vaccine. The method comprises the following steps: preparing an inactivated rabies virus ultrafiltration concentrate: purifying the inactivated rabies virus ultrafiltration concentrate by chromatography; the chromatography purification specifically comprises the following steps: column cleaning: loading a composite filler into a chromatography column, connecting an AKTA to the chromatography column, cleaning, sterilizing, and setting aside; chromatography operation: equilibrating the chromatography column with PBS buffer; sample loading: loading the inactivated rabies virus ultrafiltration concentrate into the chromatography column, eluting with an eluent, and collecting the eluate corresponding to the absorption peak at an ultraviolet wavelength of 280 nm, which is the purified inactivated rabies virus vaccine; column cleaning; and results showing that the residual protein content is as low as 5.0 ng / ml, the antigen content can reach above 16.0 IU / ml, the host protein removal rate can reach above 99%, and the antigen content recovery rate can reach above 86%.

The present invention belongs to the field of medicinal chemistry and relates to a class of 4'-fluoro-1,2,3-triazole nucleoside analogs, their preparation methods, and their use as anti-rabies virus inhibitors. The structural formula of the triazole nucleoside analogs of the present invention is: #imgabs0#;R 1 、R 2 is benzyl, 4-methoxybenzyl, 4-bromobenzyl or 2-naphthylbenzyl; R 3 The present invention relates to a method for preparing a cyclopropyl group, a phenyl group, a 1-methyl-1-hydroxyethyl group, or a methoxycarbonyl group. Most reactions in the preparation method of the present invention are insensitive to air and water, and multi-step reactions can be continued without purification, greatly simplifying the operation, reducing the difficulty of synthesis, and being suitable for large-scale preparation. Such compounds have anti-rabies virus activity and can be used to prepare anti-rabies virus inhibitors.

The present invention discloses a method and application for detecting rabies virus by an optoelectrochemical aptasensor. The aptasensor is prepared through the following steps: CdTe QDs are assembled on the surface of a carbon-doped TiO2 nanoparticle electrode by electrostatic adsorption; NH2-RNA-SH / Au NPs are immobilized on the TiO2:C-NP / CdTe electrode through a classical EDC coupling reaction between the carbonyl group on CdTe QDs and the amino group of the probe, thereby constructing the aptasensor TiO2:C-NP / CdTe / AuNPs / PRNA; then, monoethanolamine is used to block the unbound sites on the electrode surface to avoid false positives caused by non-specific binding. The present invention utilizes the efficient exciton energy transfer between cadmium telluride quantum dots and gold nanoparticles, and can quickly and reliably diagnose rabies virus, with the lowest detection concentration limit being approximately 2.6 ffu / mL.

The present application belongs to the technical field of biological medicines, and particularly relates to a mRNA vaccine against rabies viruses and a preparation method therefor, the nucleotide sequence of said vaccine being shown as any one of SEQ ID NO. 1-3. The mRNA vaccine obtained by the present application can generate an neutralizing antibody in a high level, has broad-spectrum neutralizing activity on representative strains of seven pandemic populations of rabies viruses in China, effectively induces B cell immunity and T cell immunity, and provides good protection effects for mice challenged with viruses in pre-exposure and post-exposure immune tests. In addition, the vaccine can be preserved stably for a long period under a condition of -20℃, thereby satisfying the requirements of use in countries and regions of various economic conditions.

This utility model relates to the field of biopharmaceutical technology, and in particular to a bioreactor for monitoring rabies virus cell culture. It includes a tank and a detection mechanism. The detection mechanism is located on one side of the tank, and an assembly mechanism is located at one end of the tank. In this rabies virus cell culture monitoring bioreactor, the solution is first placed inside the tank through the detection mechanism, and a top cover is fixed to the tank via a threaded connection. The detection cover is used to seal the sample to be tested. An integrated head integrates a heating rod and is electrically connected to a battery pack to power the heating rod, thereby heating the sample inside the tank. The heating rod is located inside a sensor so that the sensor can monitor the sample status (such as gas, temperature, pressure, etc.) in real time when heated. The sensor is connected to the integrated head through a connector and a socket to form a closed detection circuit. The detection signal is transmitted to a display screen via the integrated head to display the detection result. If an abnormality occurs, an alarm light flashes.

The application discloses a single-chain antibody, the amino acid sequence of which is shown as SEQ ID NO:1, and the nucleotide sequence of which is shown as SEQ ID NO:2. The single-chain antibody against rabies virus is screened from an antibody library of immunized mice by using a phage display technology. Compared with a traditional cell fusion technology, the phage display technology is simple in operation, low in cost, short in time, and high in antibody discovery and screening efficiency. The single-chain antibody obtained by the application is low in molecular weight and immunogenicity, high in antibody specificity and affinity, and has the activity of neutralizing rabies virus, thereby providing a new idea for basic research of rabies virus and development of therapeutic drugs.

This invention discloses nanobodies against rabies virus G protein and their applications, belonging to the field of nanobody technology. The nanobodies are antibody 11G6, antibody 9F7, or antibody 9A3; the amino acid sequence of antibody 11G6 is shown in SEQ ID NO. 8; the amino acid sequence of antibody 9F7 is shown in SEQ ID NO. 13; and the amino acid sequence of antibody 9A3 is shown in SEQ ID NO. 18. This invention uses phage display technology to screen for nanobodies with high affinity and high neutralizing activity. Based on this, this invention utilizes the advantage of the small size of nanobodies and verifies their unique ability to cross the blood-brain barrier (BBB) ​​through Transwell experiments, overcoming the limitation that large molecules such as antibodies and drugs cannot cross the BBB to enter the brain. This invention provides a new drug option for the treatment of rabies virus and has significant clinical implications.

Provided is an anti-rabies virus combined monoclonal antibody preparation. The present invention relates to the technical field of biomedicine. The anti-rabies virus combined monoclonal antibody preparation comprises an R92 antibody and an R71 antibody. A light chain sequence of the R92 antibody is set forth in SEQ ID NO: 1, and a heavy chain sequence of the R92 antibody is set forth in SEQ ID NO: 2. A light chain sequence of the R71 antibody is set forth in SEQ ID NO: 3, and a heavy chain sequence of the R71 antibody is set forth in SEQ ID NO: 4. The ratio of the R92 antibody to the R71 antibody in the anti-rabies virus combined monoclonal antibody preparation is (1-5):(1-5). The anti-rabies virus combined monoclonal antibody preparation has a high affinity for the rabies virus glycoprotein, has relatively good neutralizing capacity and thermal stability, and has very good neutralizing capacity against a variety of street strains. Since the two antibodies target different epitopes, the preparation can provide broader protection.

The invention discloses a recombinant rabies virus genome plasmid, a recombinant rabies virus and a preparation method and application thereof, and the preparation method specifically comprises the following steps: (1) introducing a sequence 1 and a sequence 2 into the recombinant rabies virus genome plasmid, so that the recombinant rabies virus genome plasmid has more selectivity in modification and application; (2) introducing a fluorescent protein gene and a functional probe into the recombinant rabies virus genome plasmid, so that the recombinant rabies virus genome plasmid has the functions of expressing the fluorescent protein and the functional probe; (3) preparing a corresponding recombinant rabies virus which can express fluorescent protein with high sensitivity for cell marking and can also effectively express a functional probe for functional research; the method has wide application value and market prospect in the aspects of neural circuit structure and function analysis, neuron sparse marking, vaccine and drug research and development, disease model establishment, neural network loss and reconstruction detection, virus replication or pathogenic mechanism research and the like.

This disclosure provides an attenuated suid herpesvirus 1 (a Pseudorabies virus) wherein the TK, gl and gE genes thereof are modified relative to a parent field strain, such that the resultant virus is safe and effective for use as a live vaccine that protects swine animals from challenge with a virulent Pseudorabies virus.

The invention provides a high-yield rabies virus cell line as well as a preparation method and application thereof. Specifically, the invention provides an engineered cell for producing the recombinant rabies virus, the recombinant rabies virus with effective titer can be produced by using the cell virus through one-time passage, and a neural circuit tracing tool with excellent performance is provided for the application of the defective rabies virus in neural network reverse labeling. The research in the brain science field is promoted.

The application discloses application of siRNA for targeting and inhibiting rabies virus G gene and / or L gene in preparation of a medicine for inhibiting rabies. Furthermore, the application provides siRNA-G for targeting and inhibiting rabies virus G gene and siRNA-L for targeting and inhibiting rabies virus L gene, wherein the nucleotide sequence of the sense strand of the siRNA-G is shown as SEQ ID NO:1, the nucleotide sequence of the antisense strand of the siRNA-G is shown as SEQ ID NO:2; the nucleotide sequence of the sense strand of the siRNA-L is shown as SEQ ID NO:3, and the nucleotide sequence of the antisense strand of the siRNA-L is shown as SEQ ID NO:4. The siRNA-G and / or the siRNA-L can effectively inhibit the expression of the rabies virus G gene and the L gene, reduce the proliferation of the rabies virus, and can be used as an effective small nucleic acid medicine in the development process of an anti-rabies virus medicine.