281 results about "Virus-like particle" patented technology

The invention provides a mutant of enterovirus 71 and virus-like particles thereof, and relates to the field of biological medicines. The enterovirus 71 type virus-like particle is obtained by modifying structural genes VP0, VP1 and VP3 of virus capsid protein by utilizing computational structure biology and completing self-assembly in vivo through a hansenula polymorpha expression system. Compared with the unmodified enterovirus 71 type mutant and the virus-like particles thereof, the enterovirus 71 type mutant and the virus-like particles thereof have the advantages that the immunogenicity can be obviously improved, and the clinical application prospect is relatively high.

The invention belongs to the technical field of biological pharmacy, and particularly relates to influenza virus-like particles for chimeric expression of new coronavirus RBD protein as well as a preparation method and application of the influenza virus-like particles. The virus-like particle is prepared by embedding fusion protein on the surface of influenza virus M1, and the fusion protein comprises an HA2 segment of HA protein of influenza virus, an extracellular functional region of M2e matrix protein, a conserved region short peptide of nucleoprotein, and RBD tandem protein with mutated five sites of 417, 452, 478, 484 and 501 of new coronavirus; the virus-like particles can be used for preventing specific influenza and new crowns; the preparation method of the virus-like particles is simple and low in cost, the virus-like particles are good in immunogenicity and cross protectiveness, the preparation requirements of large-scale influenza vaccines can be met, the biological safety is high, and the immunity of an organism to various influenza viruses and new coronal viruses can be stimulated.

Provided herein are SARS-CoV-2 virus-like particles as well as methods and compositions for generating SARS-CoV-2 virus-like particles. The SARS-CoV-2 virus-like particles can load and deliver transcripts (including engineered transcripts that can include therapeutic agents) into cells expressing SARS-CoV-2 entry factors. The SARS-CoV-2 virus-like particles are also useful for detecting immune response in antibodies from subjects.

The invention discloses an influenza C virus HEF protein polyclonal antibody preparation method, which comprises: carrying out signal peptide prediction, antigen epitope screening and hydrophobicity analysis through bioinformatics, constructing a recombinant prokaryotic expression vector pET32a-HEF by using a seamless cloning technology, and efficiently expressing the HEF protein in Escherichia coli to obtain the influenza C virus HEF protein polyclonal antibody. The inclusion body protein yield is improved by optimizing conditions; and purifying by nickel column affinity chromatography to obtain high-purity recombinant protein, emulsifying the high-purity recombinant protein and a Freund's adjuvant, immunizing a Japanese white rabbit, immunizing for three times, and collecting high-titer serum. The method breaks through detection limitation caused by antigen tag concealment in a eukaryotic expression system, the obtained polyclonal antibody is high in specificity, the eukaryotic expression HEF protein can be accurately recognized, and a key tool is provided for ICV virus-like particle identification, vaccine research and development and diagnosis technologies. Compared with a traditional method, the scheme is simple and convenient to operate, low in cost and high in antibody titer, and has remarkable application value.

The invention provides a sexavalent norovirus immune composition. The sexavalent norovirus immune composition comprises virus-like particles or active fragments of GI.1 type norovirus, GII.2 type norovirus, GII.3 type norovirus, GII.4 type norovirus, GII.6 type norovirus and GII.17 type norovirus. The invention also provides a kit comprising the sexavalent norovirus immune composition. The invention also provides application of the sexavalent norovirus immune composition and the kit. The composition or the kit provided by the invention can realize effective immunization aiming at six valence types of GI.1 type, GII.2 type, GII.3 type, GII.4 type, GII.6 type and GII.17 type, and can be effective for a long time after administration.

The present invention relates to a method for producing higher titers and / or a higher quality of recombinant viruses and / or less or reduced impurities in virus like particle (VLP) production in respective production processes in mammalian cell lines, the method comprising the step of supplementing a suitable mammalian cell line culture media with at least one intermediate substrate of the tricarboxylic acid (TCA) cycle and optionally supplementing the suitable mammalian cell line culture media with further additives and / or further adjusting the conditions of the mammalian cell line culture media. Furthermore, the present invention relates to the balanced suitable mammalian cell line culture media and the use of this balanced suitable mammalian cell line culture media for producing higher titers and / or a higher quality of recombinant viruses and / or less or reduced impurities in virus like particle (VLP) production in respective production processes for producing higher titers and / or quality of recombinant viruses.

The application discloses a reagent, a method and application of the reagent and the method for simultaneously detecting three common viruses in a fancy carp, and belongs to the technical field of fish quarantine. The application provides a reaction system and a detection method for simultaneously detecting three common viruses in a fancy carp, establishes a multiplex fluorescence PCR method, adopts a full-closed reaction, solves the problem that common PCR is prone to generating aerosols and causing false positive results of tests, has high detection sensitivity, can detect target genes with a copy number of 10 orders of magnitude at the minimum, is suitable for detecting a small amount of viruses carried in normal fancy carp or viruses in water quality environment, and can fully meet the requirements of customs quarantine prevention and control, has strong specificity, has no cross reaction to templates such as IHNV, RSIV and EHNV, and further provides a virus-like particle which can be used as a positive reference product in the multiplex system, is stable in state and beneficial to storage, can monitor an extraction process, and ensures that a test is established.

The present invention provides a fusion protein and an expression vector for enhancing the soluble expression of norovirus antigens. More specifically, it provides a combination of diverse genotypes of norovirus antigen proteins and RID mutations that, when norovirus antigens are used as target proteins, fold most efficiently during the formation of recombinant fusion proteins, improve the soluble expression yield of the produced fusion proteins, and enhance the assembly efficacy and homogeneity of norovirus VLPs. Furthermore, by utilizing mutational RIDs, the present invention provides a method for rapidly and in large quantities producing not only GII.4 but also other diverse genotypes of norovirus VLPs in E. coli, thereby enabling the development of VLP-based multivalent vaccines containing a wider range of genotypes.

The invention discloses akabane disease virus recombinant virus-like particles and a preparation method thereof, and belongs to the technical field of biology. The akabane disease virus recombinant virus-like particles are obtained by recombinant expression of structural protein Gn and structural protein Gc of akabane disease virus in an insect cell-baculovirus expression system and self-assembly; the amino acid sequence of the structural protein Gn is as shown in SEQ ID NO. 1; the amino acid sequence of the structural protein Gc is as shown in SEQ ID NO. 2. According to the invention, genes of coding structural proteins Gn and Gc are optimized according to codon preference of insect cells, and AKAV virus-like particles with higher safety are successfully prepared through a baculovirus-insect cell expression system. The AKAV virus-like particle provided by the invention has application potential in various fields of AKAV vaccines, immunotherapy and the like, and provides technical reserve for biological safety monitoring.

Disclosed herein are vaccine constructs for producing a virus-like particle (VLP) capable of raising an immune response to an immunogen, and uses thereof, wherein the constructs comprise nucleic acid sequences encoding an immunogen and a polyprotein, wherein the polyprotein comprises two or more viral structural proteins, wherein at least two of the two or more viral structural proteins are separated by a signal peptidase sequence such that, when the polyprotein is expressed in a host cell, the signal peptidase sequence undergoes host cell peptidase-dependent cleavage to liberate the two or more viral structural proteins, thereby allowing the liberated structural proteins to self-assemble into a VLP carrying the immunogen.

Provided are modified virus-like particles (VLPs) of paramyxoviruses, compositions containing them, methods of using the VLPs for delivery of any particular protein of interest to any of a variety of cells, kits that contain expression vectors for making, using and detecting VLPs, and methods for screening for anti-viral compounds using the VLPs. The modified VLPs contain a contiguous recombinant polypeptide that contains i) all or a segment of a C-terminal domain of a paramyxovirus nucleocapsid protein and ii) a polypeptide sequence of a distinct protein. Non-covalent complexes of paramyxovirus M protein and fusion proteins that contain a C-terminal domain of a paramyxovirus nucleocapsid protein and a polypeptide sequence of a distinct protein are provided, as are non-covalent complexes of cells, and cell receptors, with modified VLPs.

The invention discloses a virus-like particle coated with an aluminum-containing metal organic framework mineralization layer and application of the virus-like particle. In order to improve the VLPs vaccine stability and immune effect, the invention synthesizes a novel aluminum-containing metal organic framework (ZAM). The ZAM can mineralize the VLPs at a high level under a mild condition to form the VLPs-ZAM nano vaccine. Taking a foot and mouth disease virus (FMDV) virus-like particle (VLPs) vaccine as an example, a heat treatment test shows that ZAM mineralization significantly improves the heat stability of FMDVVLPs, and the effect is superior to that of Al (OH) 3 and ZIF-8. The FMDV VLPs-ZAM not only has the effect of promoting APCs to take in FMDVVLPs, but also can promote antigens to escape from lysosome to cytoplasm due to the pH responsiveness of the FMDV VLPs-ZAM. A mouse immune test shows that ZAM mineralization improves the specific immune response level and stability induced by FMDV VLPs. The invention provides a new technical means for improving the stability of the VLPs vaccine and the immune effect of the VLPs vaccine.

The present disclosure relates to virus-like particles (VLPs) comprising one or more antigens for use as vaccines. The present disclosure also relates to the use of the vaccine for the treatment of an SARS-CoV-2 infection or a coronavirus disease 2019 (COVID-19).

The invention discloses an anti-Ebola virus VP40 protein nano antibody and application of the anti-Ebola virus VP40 protein nano antibody in Ebola virus detection. The anti-Ebola virus VP40 protein nano antibody is a 3E07 antibody. The anti-Ebola virus VP40 protein nano antibody prepared by the invention has the characteristics of high sensitivity, wide binding spectrum and the like. A double-antibody sandwich enzyme-linked immunosorbent assay method constructed by the 3E07 and 3F06 antibodies can be used for efficiently detecting VP40 proteins and virus-like particles of various subtype Ebola viruses such as Zaire, Sudan, Bandong and the like, and the detection sensitivity for the VP40 proteins of the Zaire type Ebola viruses can reach 0.039 ng / mL; the antibody combination has a wide application prospect in diagnosis and pathogen detection of Ebola virus diseases.

The application discloses nanobodies against E protein of chikungunya virus and application in virus detection, and the nanobodies against E protein of chikungunya virus include N055 and 10G4 antibodies, and both have high affinity and binding activity with antigens. A double antibody sandwich enzyme-linked immunosorbent assay method constructed by the nanobodies of the N055 and 10G4 antibodies can efficiently detect E protein of chikungunya virus and virus-like particles, and the detection sensitivity can reach 49 pg / mL, and the antibody combination has wide application prospects in diagnosis and pathogen detection of chikungunya virus disease.

The invention discloses a GNRH-I-MS2 fusion protein, a castrated MS2 virus-like particle subunit vaccine and a preparation method of the castrated MS2 virus-like particle subunit vaccine.

The present invention provides: a culture medium composition which contains a pyruvate dehydrogenase kinase inhibitor and is used for promoting the production of a virus having a target gene mounted thereon when culturing a virus-producing cell into which the target gene has been introduced; a culture medium composition for promoting the production of a virus or a virus-like particle when culturing a cell that produces the virus or the virus-like particle; and the like.

The present disclosure relates to a composition comprising one or more ribonucleic acids (RNAs) encoding virus-like particle (VLP) forming elements of an influenza virus. The present disclosure further provides uses of the composition.

A vaccine for use in the prophylaxis and / or treatment of a diseaseThe present invention relates to an adenoviral vector capable of encoding a virus-like particle (VLP), said VLP displaying an inactive immune-suppressive domain (ISD). The vaccine of the invention shows an improved immune response from either of both of the response pathways initiated by CD4 T cells or CD8 T cells.

The invention relates to the field of biological medicine, and provides a coxsackie virus A16 type mutant and a virus-like particle thereof, compared with a wild type coxsackie virus A16 type, the mutant has at least one amino acid mutation site in the following regions: (1) a canyon region of a virus capsid; (2) a channel area at a secondary axis; or (3) a channel region at a quasi-tertiary axis. The mutant and the virus-like particle are obtained by optimizing and modifying structural proteins VP0, VP1 and VP3 which form a virus capsid by using computational biology and structural biology methods, co-expressing and co-assembling the structural proteins in a hansenula polymorpha expression system to form the virus-like particle, and then carrying out series of chromatographic purification. Compared with a non-mutated coxsackie virus A16 type virus-like particle mutation scheme, the coxsackie virus A16 type virus-like particle mutation scheme disclosed by the invention has the advantages that the antigen immunogenicity can be obviously improved, and the clinical application value is realized.

The invention belongs to the technical field of biological medicine and disease prevention and treatment, and particularly relates to an SSEA4 conjugated virus-like particle anti-cancer vaccine as well as a preparation method and application thereof. Specifically, according to the vaccine, an SSEA4 antigen is efficiently synthesized through a chemical enzyme method, then the SSEA4 antigen is covalently coupled with a virus-like particle (VLP), the SSEA4 antigen and the VLP are combined with a monophosphoryl lipid A adjuvant with high safety, an organism can be induced to generate strong and lasting SSEA4-resistant humoral immune response, the generated antibody can specifically recognize and kill SSEA4 positive tumor cells, and the SSEA4 positive tumor cells can be effectively killed. Meanwhile, good safety is shown in an animal model, a new solution is provided for immunotherapy of SSEA4 positive tumors, and therefore the SSEA4 positive tumor immunotherapy polypeptide has wide application prospects and market value.

Embodiments of the invention disclosed herein involve the selective engineering of Sindbis virus so as to generate cells that make enveloped virus-like particles that contain therapeutic mRNAs, and whose membrane proteins have been mutated to serve as modular binding sites for cell-targeting ligands.

The invention discloses a method for preparing a bacterial expression liquid by using hyphantria cunea V-ATPase D gene hpRNA and application of the bacterial expression liquid, an hpV-ATPase D expression vector or an MS2 + hpV-ATPase D expression vector is constructed based on a targeted hyphantria cunea V-ATPase D gene hpRNA fragment and a virus-like particle MS2 protein gene, the expression vector is introduced into a bacterial competent cell, the MS2 protein and the hpRNA of the targeted hyphantria cunea V-ATPase D gene are continuously and massively expressed through IPTG induction, and the bacterial expression liquid is obtained. According to the invention, the MS2 protein expressed by the MS2 protein gene protects hpRNA of the hyphantria cunea V-ATPase D gene from being degraded by nuclease through a wrapping effect, so that the hyphantria cunea V-ATPase D gene has good stability; tests show that both the hpV-ATPase D and the MS2 + hpV-ATPase D have an efficient fatality rate on fall webworms and have a remarkable inhibition effect on development and reproduction of the fall webworms. The bacterial expression liquid containing the hyphantria cunea V-ATPase D gene hpRNA is used for preventing and treating hyphantria cunea, has the characteristics of high practicability, convenience, rapidness, high efficiency and sensitivity, and provides a theoretical basis for researching a hyphantria cunea prevention and treatment method.

The present invention discloses a recombinant virus-like particle of Akabane disease virus and a preparation method thereof, and belongs to the field of biotechnology. The recombinant virus-like particle of Akabane disease virus is obtained by recombinantly expressing the structural protein Gn and the structural protein Gc of Akabane disease virus in an insect cell-baculovirus expression system and self-assembly; the amino acid sequence of the structural protein Gn is shown in SEQ ID NO.1; the amino acid sequence of the structural protein Gc is shown in SEQ ID NO.2. The present invention optimizes the genes encoding the structural proteins Gn and Gc according to the codon preference of insect cells, and successfully prepares safer AKAV virus-like particles through the baculovirus-insect cell expression system. The AKAV virus-like particles provided by the present invention have application potential in various fields such as AKAV vaccines and immunotherapy, and provide technical reserves for biosafety monitoring.

The disclosure relates to a truncated recombinant L1 protein of Human papillomavirus (HPV), a recombinant polynucleotide encoding the protein, a recombinant vector comprising the polynucleotide, an isolated cell comprising the recombinant vector, a composition comprising the protein, and a method for producing the protein. The disclosure also relates to an HPV virus-like particle (VLP) comprising the protein, an immunogenic composition comprising the protein or the VLP, and a method for inducing an immune response to HPV in an individual.

The present disclosure provides virus-like particles (VLPs) comprising: i) a CRISPR-Cas effector polypeptide; ii) a recombinant lentivirus comprising a nucleotide sequence encoding a therapeutic polypeptide having a length of from about 250 amino acids to about 3,000 amino acids, where the VLP comprises a pseudotyping viral glycoprotein and / or a polypeptide that provides for binding to a target cell. The present disclosure provides systems for producing a VLP. The present disclosure provides methods of delivering a therapeutic protein, using a VLP of the present disclosure.

The disclosure provides an animal cell stably expressing a virus-like particle (VLP). The disclosure also provides a method for manufacturing a virus-like particle, a virus-like particle, a vaccine composition, a method for preventing viral infection, and a method for producing antibodies.

The invention discloses a method for analyzing related impurities in a virus-like particle drug, which adopts a tandem chromatography system consisting of a large-aperture size exclusion chromatographic column A and a small-aperture composite chromatography chromatographic column B which are directly connected, the column B has a separation mechanism of volume exclusion and hydrophobic interaction, chromatographic signals are detected, and the related impurities in the virus-like particle drug are analyzed. Synchronous separation, identification and quantification of virus-like particle related components, capsid protein related components and free oligonucleotide related components are realized by analyzing the peak appearance sequence of each chromatographic peak and the absorption signal intensity ratio under different wavelengths. The method effectively solves the problem of co-elution of different types of impurities, especially capsid proteins and free oligonucleotides in a traditional chromatographic method, has the advantages of high separation degree, accurate quantification and good repeatability, and is suitable for process development and quality control of virus-like particle drugs.

The application discloses a virus-like particle vaccine freeze-drying protective agent and a preparation method thereof. The freeze-drying protective agent comprises A liquid and B liquid mixed in a volume ratio of 1:1 to 3:1; the A liquid comprises the following components with terminal concentrations: 0.01-0.5 mol / L base liquid I, 0.5-2 mmol / L base liquid II, 0.01-0.1% polysorbate 80, 5-20% sucrose and 1-5% trypsin; and the B liquid comprises the following components with terminal concentrations: 0.1-5% trehalose, 0.1-5% oligogalactose and 0.1-2% PEG6000. Through the compounding of the A liquid and the B liquid, the application can effectively enhance the immunization effect of the vaccine, prolong the storage time of the vaccine and keep the virus-like particle form intact.

The invention relates to an extracellular vesicle (EV), viral particle or virus like particle (VLP) comprising an RNA molecule attached to the internal surface of the EV membrane, to the viral particle or to the VLP membrane via a fusion protein comprising an EV membrane, viral particle or VLP protein or part thereof and an RNA-binding domain, wherein the RNA molecule comprises at least one stemloop, an siRNA sequence and a sequence that is recognized by the RNA-binding domain, and wherein the sequence that is recognized by the RNA-binding domain is bound by the RNA-binding domain, to encoding nucleic acid molecules and uses thereof.