250 results about "Viral protein" patented technology

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.

The application belongs to the technical field of immunology and in vitro diagnosis, and particularly relates to a rabbit-derived recombinant monoclonal antibody specifically recognizing type II grass carp reovirus VP4 protein, a eukaryotic expression method and application. The S6 gene in GCRV-II encodes VP4 protein, the application transfects the target gene S6 into HEK293 cells for expression and purification, cooperates with Freund's adjuvant to immunize New Zealand white rabbits, and uses ELISA, single B cell screening and eukaryotic expression technology to obtain a rabbit-derived recombinant monoclonal antibody specifically recognizing GCRV-II VP4 protein. The antibody has strong specificity, provides support for further establishment of specific type II grass carp reovirus diagnosis technology, and has great application value for development of GCRV-II related scientific research.

The invention provides a porcine deltacoronavirus S1 protein nanoparticle vaccine CTDnps and application thereof, and belongs to the technical field of vaccine preparation. The PDCoV S1 protein C-terminal structural domain nanoparticle vaccine capable of being self-assembled, provided by the invention, comprises ferritin and a PDCoV S1 protein C-terminal structural domain, the amino acid sequence of the ferritin is as shown in SEQ ID NO.1, and the amino acid sequence of the PDCoV S1 protein C-terminal structural domain is as shown in SEQ ID NO.2. The PDCoV S1 protein C-terminal structural domain nanoparticle vaccine capable of being self-assembled comprises the ferritin and the PDCoV S1 protein C-terminal structural domain. The PDCoV S1 protein C-terminal structural domain nanoparticle vaccine (S1CTD-Fer) constructed by the invention can be self-assembled to form ferritin-like cage-shaped nanoparticles, and the hydrodynamic diameter of the cage-shaped nanoparticles is obviously greater than that of natural ferritin nanoparticles. Compared with the S1CTD, the S1CTD-Fel based on the ferritin carrier can obviously enhance the immunogenicity of the S1CTD, can obviously enhance the uptake efficiency of antigen presenting cells, and is high in safety.

The invention discloses a recombinant yeast for expressing large yellow croaker iridovirus MCP protein and a preparation method and application of the recombinant yeast in preparation of vaccines, and is characterized in that the recombinant yeast is saccharomyces cerevisiae transformed with pYD1-MCP recombinant plasmids and is named as EBY100 / pYD1-MCP, the pYD1-MCP recombinant plasmids can express the MCP protein on a yeast surface display system, the nucleotide sequence of a gene segment of the MCP protein is shown as SEQ ID NO.1, the nucleotide sequence of the gene segment of the MCP protein is shown as SEQ ID NO.2, and the nucleotide sequence of the gene segment of the MCP protein is shown as SEQ ID NO.2. The preparation method comprises a step of constructing a recombinant plasmid pYD1-MCP and a step of constructing a recombinant yeast EBY100 / pYD1-MCP, the oral vaccine of the larimichthys crocea iridovirus vaccine can be prepared, and the oral vaccine has the advantages of being high in protection rate, safe, low in cost, convenient to use and good in application prospect in prevention and control of the larimichthys crocea iridovirus.

The invention discloses a liquid-phase protein chip method for simultaneously detecting three kinds of porcine coronaviruses and a kit thereof, which comprises the following steps of: after activating and coupling microspheres, respectively coating antigen on porcine delta coronavirus N protein, porcine acute diarrhea syndrome coronavirus N protein and porcine epidemic diarrhea virus N protein by using the microspheres; matching the two fluorescent dyes according to different concentrations to form fluorescent coding microsphere combination arrays with different energy levels; the microspheres are analyzed one by one based on the flow principle, red laser recognizes microsphere codes, green laser reads fluorescence intensity of report molecules, computer analysis and processing are carried out, and fluorescence signals are converted into quantitative detection results; and determining the optimization of the detection conditions of the liquid-phase protein chip after the liquid-phase protein chip test. The antigens coated on the microspheres only specifically react with corresponding antibodies and are not combined with antibodies generated by other pathogens; the PDCoV, SADS-CoV and PEDV antibodies in the sample to be detected can be rapidly, effectively and accurately detected, and reference is provided for prevention of the porcine coronavirus.

A construction and use of an environmentally adaptive co-regulated mRNA nanodelivery system. By incorporating EACR molecules into mRNA nanoparticles, the microenvironment is remodeled to be suitable for robust and sustained mRNA expression, while tissue damage associated with the self-immunogenicity of mRNA drugs is avoided. The nanodelivery system is compatible with ionizable lipid nanoparticles, cationic liposomes, cationic nanoemulsions, and polymeric nanoparticles. The EACR molecules include: anti-inflammatory drugs, tyrosine kinases / adaptors, JAK / STAT and MAPK pathway inhibitors, nutrients and metabolites, membrane transporters / ion channels, phosphodiesterase and cellular stress inhibitors, and natural viral proteins. The therapeutic mRNAs may encode tumor, viral, or bacterial antigens, immunomodulatory factors, therapeutic antibodies, or functional proteins / enzymes, and can play a role in the fields of regenerative medicine, protein supplementation / replacement therapy, targeted gene editing, and immunotherapy.

The invention belongs to the technical field of biology, and particularly relates to a swine monoclonal antibody of hog cholera virus Erns protein. The amino acid sequence of a heavy chain variable region of the swine monoclonal antibody is shown as SEQ ID NO: 1, the amino acid sequence of a heavy chain constant region of the swine monoclonal antibody is shown as SEQ ID NO: 2, the amino acid sequence of a light chain variable region of the swine monoclonal antibody is shown as SEQ ID NO: 3, and the amino acid sequence of the light chain constant region of the swine monoclonal antibody is shown as SEQ ID NO: 4. The full-length genes of the heavy chain and the light chain of the swine monoclonal antibody are obtained from a swine B cell specifically amplified by the same hog cholera virus Erns protein, the natural structure of an antibody molecule is reserved, affinity loss possibly caused by artificial recombination is avoided, and the swine monoclonal antibody has high affinity and specificity, and can be used for preparing the swine monoclonal antibody. And a precise molecular tool is provided for antigen structure analysis of the hog cholera virus Erns protein and establishment of a hog cholera diagnosis method. The monoclonal antibody is applied to a blocking ELISA detection method of the hog cholera virus Erns antibody, and the established Erns antibody blocking ELISA detection method is high in sensitivity and strong in specificity.

This invention belongs to the field of biodetection technology, specifically relating to a monoclonal antibody 3C7 for detecting BK virus VP1 protein and its applications. The amino acid sequences of the heavy chain variable regions CDR1, CDR2, and CDR3 of the monoclonal antibody are SEQ ID NO. 1–3, and the amino acid sequences of the light chain variable regions CDR1, CDR2, and CDR3 are SEQ ID NO. 4–6, respectively. This antibody exhibits high specificity and can recognize the BK virus VP1 protein. The test strip includes a backing plate, a sample pad, a colloidal gold pad, a nitrocellulose membrane, and absorbent paper. The detection method provided by this invention is simple to operate, highly sensitive, and requires no instruments, enabling rapid and visual detection of BK virus VP1 in urine or recombinant protein samples. It is suitable for on-site screening and point-of-care testing and can supplement existing detection methods such as nucleic acid testing.

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 belongs to the technical field of antibody preparation, and particularly relates to a recombinant rabbit monoclonal antibody for specifically recognizing type II grass carp reovirus VP56 protein and application of the recombinant rabbit monoclonal antibody. The amino acid sequence of a light chain variable region of the monoclonal antibody is as shown in SEQ ID NO.3, and the amino acid sequence of a heavy chain variable region of the monoclonal antibody is as shown in SEQ ID NO.1. The recombinant rabbit monoclonal antibody for resisting the II type grass carp reovirus VP56 protein prepared by the invention has relatively strong specificity and titer, can specifically recognize the virus VP56 protein in tissues and cells in a natural state, has high affinity for recognizing and combining the virus VP56 protein, has strong anti-interference capability, and can be used for preparing the recombinant rabbit monoclonal antibody for resisting the II type grass carp reovirus VP56 protein. The method is suitable for establishing a high-specificity, high-sensitivity and high-accuracy type II grass carp reovirus diagnosis technology.

The invention belongs to the technical field of biological genetic engineering, and particularly relates to a mouse model construction scheme capable of reproducing a key phenotype of hemophagocytic syndrome, which comprises the following steps: inserting a loxP-STOP-loxP-LMP1 conditional expression cassette into a Rosa26 safe port site in a fixed point manner to obtain an Lmp1cKI mouse strain; and hybridizing the mouse with an Ncr1-Cre transgenic mouse of NK cell specific expression Cre recombinase. In a double transgenic offspring (Lmp1Ncr1-Cre), STOP fragments are cut off by Cre enzyme, and continuous high expression of EB virus LMP1 protein in NK cells is driven. The model does not need chemical induction or immune intervention, and typical hemophagocytic syndrome phenotypes, namely serum IL-6, TNF-alpha and IFN-gamma, spontaneously appear within 7-9 days after young rats are born; the body weight is reduced and the survival rate is obviously reduced; the proportion of lymphocytes is sharply reduced; and NK cells are depleted. The invention provides a stable and reliable tool for hemophagocytic syndrome pathogenesis research, drug screening and treatment evaluation.

The invention relates to a recombinant chimeric antigen aiming at bovine nodular skin disease virus, an immunogenic composition containing the recombinant chimeric antigen and application of the recombinant chimeric antigen. The bovine nodular skin disease virus recombinant chimeric antigen comprises two immunogens which are connected in series and fused according to a specific sequence: bovine nodular skin disease virus ORF122 and ORF060 proteins or antigenic fragments thereof, and can excite immune response aiming at two infectious virus particles, namely, intracellular mature virus particles (IMV) and cell outer enveloped virus particles (EEV); therefore, the specific immune protection effect on the bovine nodular skin disease virus is efficiently excited; in addition, the bovine nodular skin disease virus vaccine disclosed by the invention also has good safety, quick responsiveness and productivity support, and has an extremely good clinical application prospect.

The invention provides modified an isolated T lymphocytes having reduced or eliminated expression of the T Cell Receptor, due to reduced or eliminated expression of a CD3ζ, T Cell Receptor Alpha Chain, or T Cell Receptor Beta Chain gene relative to a T lymphocyte without modification, where the isolated T lymphocyte expresses a protein comprising US6, UL40 viral protein and signal peptide, and UL18.

Provided herein is a nucleic acid encoding a prion protein, wherein the prion protein (a) comprises an amino acid sequence comprising a protective sequence variant such as a G127V mutation and (b) lacks a fully functional 3' glycosylphosphatidylinositol (GPI)-anchor attachment sequence. Also provided are viral vectors and pharmaceutical compositions comprising the nucleic acids. Further provided are methods and uses of treating prion diseases or other neurodegenerative diseases that confer toxicity through cell surface- anchored prion proteins in a subject, comprising administering to the subject an effective amount of a pharmaceutical composition or viral vector described herein.

The invention provides a human monoclonal antibody B1AG for resisting type 4 adenovirus. The neutralizing antibody for resisting the 4-type adenovirus provided by the invention can be specifically combined with the 4-type adenovirus, and has very high affinity activity and very strong neutralizing activity at low concentration. The invention also provides application of the antibody in preparation of medicines for treating and / or preventing type 4 adenovirus diseases, and a detection kit containing the antibody.

The application discloses a tandem antigen epitope peptide of G protein of Paralichthys olivaceus rhabdovirus and application thereof, and belongs to the field of fish molecular immunology. The amino acid sequence of the tandem antigen epitope peptide is shown as SEQ ID NO:1. The preparation method of the application comprises the following steps: (1) first, analyzing the structural characteristics of HIRRV-G protein and predicting B cell antigen epitopes, and synthesizing the antigen epitopes with advantages based on the prediction results; (2) then, screening candidate peptide segments with high affinity through enzyme-linked immunosorbent assay; (3) sequentially connecting the high-affinity peptide segment sequences meeting the requirements by using a GPGPG linker, and cloning the connected sequences into a pET-28a prokaryotic expression vector, so that the tandem antigen epitope peptide is obtained after induced expression. Compared with the full-length G protein, the tandem antigen epitope peptide has smaller molecular weight, stronger stability and stronger hydrophilicity; the tandem antigen epitope peptide can induce a large amount of specific antibodies in fish bodies, and significantly reduces the mortality of fish infected by the virus. The tandem antigen epitope peptide can be used for the development of HIRRV diagnostic reagents and subunit vaccines.

The invention discloses a monoclonal antibody 5B11 for recognizing an extracellular domain of peste des petits ruminants virus H protein and application of the monoclonal antibody 5B11, and belongs to the technical field of monoclonal antibodies. According to the invention, PPRV-H-185 protein with an amino acid sequence as shown in SEQ ID NO.1 is used for immunizing a mouse, a monoclonal antibody 5B11 with a neutralizing effect is prepared, a heavy chain variable region of the monoclonal antibody 5B11 comprises CDR1-3 with an amino acid sequence as shown in SEQ ID NO.7-9, and a light chain variable region of the monoclonal antibody 5B11 comprises CDR1-3 with an amino acid sequence as shown in SEQ ID NO.12-14. Experimental results show that the monoclonal antibody can be combined with a virus, so that the virus loses the ability of combining an epithelial cell H protein receptor Nectin-4 and a lymphocyte H protein receptor SLAM, and the virus is prevented from invading cells. The invention provides a new biological material for detection and prevention of peste des petits ruminants virus, and has a good clinical application prospect.

The invention belongs to the technical field of biological medicine, and particularly relates to a rabies virus G protein carrier, a rabies virus vaccine and a preparation method and application of the rabies virus G protein carrier and the rabies virus vaccine. The rabies virus G protein vector pscAAV-G is obtained by inserting a coding gene of rabies virus G protein into a coding region between inverted terminal repetitive sequences at two ends of a double-chain adeno-associated virus vector, so that the problem of expression delay of single-chain AAV (ssAAV) is solved, and rapid and efficient expression of the rabies virus G protein is realized; furthermore, an endogenous nucleic acid adjuvant is embedded into a non-coding region of the pscAAV-G, so that the rabies virus G protein carrier pscAAV-G-Adj is prepared, and Th1 type immune response is remarkably improved. According to a rabies virus vaccine prepared by using the rabies virus G protein carrier, the antigen expression efficiency and immunogenicity of an AAV carrier vaccine are remarkably improved, long-acting protection of single inoculation is realized, and the defects that an existing rabies vaccine is short in immune cycle and needs to be inoculated for multiple times are overcome.

This invention discloses methanogenic lyases with the same protein family annotation as PeiR lyases and their applications. These enzymes are obtained through homology screening based on the known characteristics of PeiR lyases targeting peptide bonds in the methanogenic cell wall, using the Pfam protein family database annotation. PeiR lyase proteins participate in the biological process of hydrolyzing archaea cell walls, effectively killing methanogens and reducing methane production. Proteins homologous to this protein have potential methane-reducing potential. This invention integrates rumen microbial metagenomic sequencing data, uses a series of bioinformatics software to screen methanogenic viral proteins, and combines Pfam functional domain annotation for homology analysis, ultimately identifying a series of lyases. These lyases were successfully expressed in a prokaryotic expression system, and in vitro gas production experiments also showed that the crude enzyme solution significantly reduced methane production.

Methods, systems, and apparatus, including computer programs encoded on a computer storage medium, for computationally docking a small molecule or ligand to a prion-protein In one aspect, a system comprises receiving data comprising a structure of a candidate drug, a structure of a prion-protein filament comprising one or more prions, and a location of a binding site in the prion-protein filament structure, generating a plurality of stacked pose configurations of the candidate drug using at least the candidate drug structure, determining a corresponding docking score for each stacked pose configuration in the plurality of stacked pose configurations in accordance with a measure of a likelihood of binding for the stacked pose configuration in the location of the binding site in the prion-protein filament, and taking an action based on the corresponding docking scores.

The application provides an anti-Spike protein antibody of a novel coronavirus and an application thereof. By means of genetic engineering and phage surface display library technology, specific antibodies against the S protein of the novel coronavirus are screened from a single-chain antibody library of non-immune full human sequences. The antibodies have an affinity to the S protein of the virus between 1nM and 50nM, the antibodies have an inhibiting effect on the combination of the S protein of the novel coronavirus and human receptor ACE2, and the anti-S protein antibodies of the application have a good ability to combine with the S protein and a potential neutralizing inhibiting effect. The application provides specific antibody candidate molecules for the development of diagnostic reagents, preventive and therapeutic antibody drugs against the novel coronavirus (2019-nCoV) and the treatment of other diseases such as pneumonia caused by the coronavirus.

The present invention discloses a monoclonal antibody capable of recognizing a novel duck reovirus NDRVσC protein, a preparation method and an application thereof, wherein the monoclonal antibody comprises a heavy chain variable region and a light chain variable region; the heavy chain variable region comprises a V sequence as shown in SEQ ID NOs: 3, 4, and 5. H CDR1, V H CDR2 and V H CDR3, the light chain variable region includes the amino acid sequences shown in SEQ ID NOs: 6, 7, and 8. L CDR1, V L CDR2 and V L CDR3. The present invention also discloses a method for preparing a monoclonal antibody and a double-antibody sandwich ELISA method based on the antibody and the σC rabbit polyclonal antibody. This method can be used to quantify NDRV subunit vaccine antigens and determine animal immunization doses. Compared with the methods reported in the literature, this ELISA method is simpler to operate and has great potential for practical application. The present invention provides a new method for evaluating NDRV immune efficacy and controlling vaccine quality, which can ensure the upgrading of NDRV vaccines.

The present invention relates to synthetic designs or chimeric proteins for pseudotyping baculovirus (Autographa californica nuclear polyhedrosis virus) with the rabies virus glycoprotein G (gG) on its surface (Bac::gG-FL), which can be used in anti-rabies vaccine formulations. The chimeric protein is designed from a gene cassette containing gene fragments of the ectodomain of the G glycoprotein of the Pasteur strain rabies virus, a linker of 7 amino acids (GGGGSGG), as well as transmembrane (TM) and cytoplasmic (CT) regions of the gp64 baculovirus protein, with the arrangement of the sequences in the designed gene cassette being shown in figure 1.

The invention provides a fully humanized monoclonal antibody for resisting SARS-CoV and SARS-CoV-2 and application of the fully humanized monoclonal antibody, and belongs to the technical field of immunology, and the amino acid sequence of a heavy chain variable region of the monoclonal antibody is as shown in SEQ ID NO: 1; the amino acid sequence of a light chain variable region of the monoclonal antibody SARS2-1 is as shown in SEQ ID NO: 2; the antibody has a unique CDR partition, can identify S proteins of the two viruses at the same time, and has cross-neutralization activity on pseudoviruses of the two viruses. The antibody has the characteristics of high expression, full human source and good stability, is suitable for industrial production, and has potential application value for coping with outbreak prevalence caused by coronavirus possibly occurring at present and in the future.

The invention relates to an infectious clone of tea tree reticulate spot virus and a method for constructing a VIGS vector thereof, and its application, which belong to the field of genetic engineering technology. The infectious clone comprises three full-length cDNA recombinant plasmids of tea tree reticulate spot virus (TPLPV), namely pTPLPV1, pTPLPV1 and pTPLPV3. pTPLPV3 can be used to insert exogenous gene fragments to form a VIGS vector of TPLPV. By Agrobacterium-mediated method, pTPLPV3-VIGS is inoculated alone or mixed with the first two chains for infection. The NbPDS gene of the model plant tobacco can be silenced and an albino silent phenotype can be produced. Infection alone can silence the CsPDS gene of the tea tree and produce an albino silent phenotype. The present invention transforms the tea tree virus TPLPV into an infectious clone and a VIGS vector for the first time, providing a tool carrier and technical support for studying the function of the viral protein, the pathogenic mechanism and the construction and transformation of the silencing vector.

The invention belongs to the field of biology, and particularly relates to construction of a recombinant Seneca virus strain with Seneca virus 2B protein translation block site mutation. The 26th lysine (K) of the 2B protein is found to be a key site for inhibiting the translation of the host protein for the first time, and after the 26th K of the 2B protein is mutated into the alanine (A), the translation retardation phenomenon of the host protein is obviously weakened; secondly, the 26th K of the 2B protein is mutated into A to construct infectious clone, and the recombinant Seneca virus strain rSVA-2B K26A with 2B protein translation retardation site mutation is successfully rescued and obtained, compared with wild type SVA, after the rSVA-2B K26A infects host cells, the translation retardation phenomenon of the host cells is weakened, the expression level of host antiviral inflammatory cytokines is enhanced, and the expression level of the recombinant Seneca virus strain rSVA-2B K26A is improved. The Seneca virus vaccine strain can be used as a potential vaccine candidate strain, and a foundation is laid for developing a novel prevention and control strategy aiming at the Seneca virus.

The application provides an anti-Spike protein antibody of a novel coronavirus and an application thereof. By means of genetic engineering and phage surface display library technology, specific antibodies against the S protein of the novel coronavirus are screened from a single-chain antibody library of non-immune full human sequences. The antibodies have an affinity to the S protein of the virus between 1nM and 50nM, the antibodies have an inhibiting effect on the combination of the S protein of the novel coronavirus and human receptor ACE2, and the anti-S protein antibodies of the application have a good ability to combine with the S protein and a potential neutralizing inhibiting effect. The application provides specific antibody candidate molecules for the development of diagnostic reagents, preventive and therapeutic antibody drugs against the novel coronavirus (2019-nCoV) and the treatment of other diseases such as pneumonia caused by the coronavirus.

The present invention relates to the production of modified influenza vial proteins in plants. More specifically, the present invention relates to producing and increasing influenza virus-like particle (VLP) production in plants, wherein the VLPs comprise the modified influenza viral proteins, such as modified influenza hemagglutinin (HA). The HA protein may comprising an amino acid sequence comprising at least one substitution when compared to a corresponding wildtype amino acid sequence. Further provided are nucleic acid encoding the modified HA protein. Furthermore methods of producing an influenza virus like particle (VLP) and methods of increasing yield of production of an influenza virus like particle (VLP) in a plant, portion of a plant, or a plant cell, are also provided.

The invention discloses a bovine nodular skin disease virus fusion antigen as well as a product and application thereof, and belongs to the technical field of immunity. In order to solve the technical problems that the immune antigen of the bovine nodular skin disease virus is poor in safety, not easy to prepare and low in immunogenicity, the technical scheme is characterized in that the fusion antigen aiming at the bovine nodular skin disease virus is provided, and the fusion antigen comprises three antigen peptide fragments derived from bovine nodular skin disease virus LSDV060 protein and LSDV122 protein, and the three antigen peptide fragments are fused through head-to-tail splicing.