106 results about "Rabies virus" patented technology

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.

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.

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 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 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.

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.

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.

The invention relates to a recombinant rabies virus. The recombinant rabies virus is obtained by inserting an antigen gene of feline calicivirus and an antigen gene of feline herpes virus into a genome of a rabies virus attenuated strain. The triple inactivated vaccine for preventing and treating the feline calicivirus disease, the feline herpes virus disease and / or the rabies is prepared based on the recombinant rabies virus, and after the triple inactivated vaccine is used for immunizing cats, neutralizing antibodies aiming at the feline calicivirus disease, the feline herpes virus disease and the rabies virus can be effectively induced and generated; the simultaneous immunization on the three viruses is realized.

The present invention discloses a rabies virus serum-free Vero cell inactivated vaccine and a preparation method thereof, and belongs to the technical field of vaccine manufacturing. The preparation method of the inactivated vaccine comprises the following steps: resuscitating Vero cells, performing serum-free adaptation and domestication of the Vero cells, preparing modified microcarriers, adding the modified microcarriers to a bioreactor, adding serum-free culture medium, inoculating the domesticated Vero cells into the bioreactor, amplifying and harvesting the cells, inoculating rabies virus, adding serum-free culture medium for virus culture, harvesting, and obtaining a virus liquid; finally, inactivating the virus, incubating, concentrating, purifying, and freeze-drying to obtain a rabies virus serum-free Vero cell inactivated vaccine. The virus liquid prepared by the present invention has the advantages of high virus content, stable production process, and low production cost, and the prepared vaccine has the effects of improving safety in use and reducing side effects.

The invention discloses an anti-rabies virus G protein nano antibody and application thereof, and belongs to the technical field of nano antibodies. The nano antibody is an antibody 11G6, an antibody 9F7 or an antibody 9A3; the amino acid sequence of the antibody 11G6 is as shown in SEQ ID NO. 8; the amino acid sequence of the antibody 9F7 is as shown in SEQ ID NO. 13; the amino acid sequence of the antibody 9A3 is as shown in SEQ ID NO. 18. According to the invention, the nano antibody with high affinity and high neutralizing activity is screened by a phage display technology. On the basis, the advantage that the size of the nano antibody is small is utilized, the unique ability of the nano antibody to penetrate through the BBB is verified through a Transwell experiment, and the defect that macromolecules such as antibodies and drugs cannot penetrate through the BBB to enter the brain is overcome. The invention provides a new drug choice for the treatment of rabies virus, and has important clinical significance.

The invention discloses an anti-rabies virus G protein monoclonal antibody, a detection reagent and application of the anti-rabies virus G protein monoclonal antibody and the detection reagent. The anti-rabies virus G protein monoclonal antibody 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 provided by the invention has high affinity and detection sensitivity to rabies virus G protein, and lays a foundation for research, development and popularization of colloidal gold test strips and fluorescence immunoassay test strips.

The present application relates to the technical field of biotechnology, in particular to a pseudorabies immunogen composition and application thereof. The present application provides a pseudorabies immunogen composition, which comprises at least one antigen saRNA (self-replicating RNA) molecule; the antigen saRNA molecule comprises at least one open reading frame (ORF), and the open reading frame ORF encodes a polypeptide comprising a gD antigen from a pseudorabies virus. The present application uses the self-replicating RNA technology to prepare a pseudorabies vaccine, and can protect the infection of the pseudorabies virus, which effectively solves the shortcoming of low protection efficiency of the previous pseudorabies vaccine on the pseudorabies virus.

The invention discloses a CDR region of a nano antibody for targeting rabies virus. The invention also discloses a nano antibody aRV7, which comprises the CDR region of the nano antibody targeting the rabies virus. The invention also discloses an antibody targeting the rabies virus. The invention also discloses a polynucleotide, an expression vector, a host cell and an antibody conjugate. The invention also discloses a CDR region of the rabies virus-targeted nano antibody, a nano antibody aRV7, the rabies virus-targeted antibody, and an application of the antibody conjugate. The invention also discloses a pharmaceutical composition. The nano antibody aRV7 and the antibody targeting the rabies virus have specific recognition and binding performance on rabies virus G protein, can target the rabies virus, and can be used for preparing reagents, kits, vaccines or drugs for detecting, diagnosing, treating and / or preventing rabies virus infection.

The invention discloses a CDR region of a nano antibody for targeting rabies virus. The invention also discloses a nano antibody aRV15, which comprises the CDR region of the nano antibody targeting the rabies virus. The invention also discloses an antibody targeting the rabies virus. The invention also discloses a polynucleotide, an expression vector, a host cell and an antibody conjugate. The invention also discloses a CDR region of the rabies virus-targeted nano antibody, a nano antibody aRV15, the rabies virus-targeted antibody, and an application of the antibody conjugate. The invention also discloses a pharmaceutical composition. The nano antibody aRV15 and the antibody targeting the rabies virus have specific recognition and binding performance on rabies virus G protein, can target the rabies virus, and can be used for preparing reagents, kits, vaccines or drugs for detecting, diagnosing, treating and / or preventing rabies virus infection.

The invention provides a broad-spectrum anti-rabies virus fully-humanized monoclonal antibody and application thereof, the fully-humanized 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: GFTFSDYE, ISSSGATK and ARVPGQQVGVDR, and the light chain variable region has three complementary determining region amino acid sequences: GFTFSDYE, ISSSGATK and ARVPGQQVGVDR. The amino acid sequences of three complementary determining regions of the light chain variable region are RAIHNY, ATS and QQYYGYPWT respectively. The fully human monoclonal antibody R06 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.

The invention relates to the technical field of rabies antibodies, in particular to an anti-RABV antibody with a neutralizing binding site at a non-epitope III and application of the anti-RABV antibody. A heavy chain variable region of the anti-RABV antibody comprises CDR1 (SEQ ID NO 2, allowed mutation of 12% or below), CDR2 (SEQ ID NO 3, allowed mutation of 12% or below) and CDR3 (SEQ ID NO 4, allowed mutation of 12% or below), and a light chain variable region of the anti-RABV antibody comprises CDR1 (SEQ ID NO 6, allowed mutation of 12% or below), CDR2 (SEQ ID NO 7, allowed mutation of 12% or below) and CDR3 (SEQ ID NO 8, allowed mutation of 12% or below). The anti-RABV antibody provided by the invention has good neutralizing activity on the rabies virus CVS-11 and related mutant strains.

The invention discloses an anti-rabies virus mutant antibody as well as a preparation method and application thereof, and belongs to the field of antibodies. The technical problem to be solved is to provide an anti-rabies virus mutant antibody with longer in-vivo half-life period and longer effective antibody concentration maintenance time. According to the key point of the technical scheme, the anti-rabies virus mutant antibody comprises a heavy chain and a light chain, and the nucleotide sequence of the heavy chain is as shown in SEQ ID NO: 1. According to the invention, a YTE mutation technology is adopted to carry out long-acting modification on the recombinant human anti-rabies virus monoclonal antibody R8. Through synthesis of heavy chain and light chain gene sequences of the mutated antibody, a mutation expression plasmid is constructed, and pharmacokinetic verification is carried out after transient transfection expression purification, so that long-acting transformation and preparation of the R8 antibody are realized, and technical support is provided for clinical application.

The application belongs to the technical field of biological medicine, and provides a photosensitizer, a targeted delivery nano platform and application. A preparation method of the photosensitizer comprises the following process: a Suzuki coupling reaction is used to synthesize an intermediate with an aldehyde group by taking a brominated or iodinated triphenylamine as a precursor and reacting with boric acid, and then a Knoevenagel condensation reaction is used to convert the terminal aldehyde group into a strong electron-withdrawing dicyanovinyl group by reacting the aldehyde group intermediate with malononitrile, so as to synthesize the photosensitizer PSs. The application takes a structure-optimized photosensitizer (used for generating ROS and inactivating viruses) and a STING agonist (used for activating the STING pathway and inducing cell death) as core treatment components, and takes a rabies virus glycoprotein peptide as a active targeting ligand, so that the "PDT-immune" synergistic inactivation of RABV can be realized.

The invention relates to a cell line for saving rabies virus and application thereof. A cell comprising, expressing and / or secreting B19G or a variant having at least 80% sequence identity thereto and retaining function is provided. According to the cell line, viruses can be efficiently rescued, and passage of the viruses is reduced.

The application discloses application of lapatinib in preparation of a rabies virus resisting medicine, and first discovers the rabies virus resisting activity of lapatinib, breaks the limitation of lapatinib only being used for treating tumors, and first proves the role of lapatinib in inhibiting rabies virus replication through in-vivo and in-vitro experiments, develops the potential application value of lapatinib in early treatment after rabies virus exposure, and provides a new use of lapatinib, i.e., using lapatinib as a candidate medicine for rabies treatment, especially specific treatment in the early stage of rabies virus infection, and fills the blank of specific treatment medicines after rabies virus exposure.