30 results about "Nucleocapsid Proteins" patented technology

The invention discloses an antibody specifically bound with novel coronavirus nucleocapsid protein or an antigen binding fragment thereof and application thereof. The antibody or the antigen binding fragment thereof comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprises a CDR1 of which the amino acid sequence is shown as SEQ ID NO: 5, a CDR2 of which the amino acid sequence is shown as SEQ ID NO: 6 and a CDR3 of which the amino acid sequence is shown as SEQ ID NO: 7; the light chain variable region comprises CDR1 of which the amino acid sequence is as shown in SEQ ID NO: 8, CDR2 of which the amino acid sequence is as shown in SEQ ID NO: 9 and CDR3 of which the amino acid sequence is as shown in SEQ ID NO: 10. The antibody provided by the scheme of the invention can specifically recognize the novel coronavirus nucleocapsid protein, and has the capability of inhibiting SARS-CoV-2 N protein from inducing excessive complement activation.

The invention relates to the technical field related to immunological detection, in particular to a mouse anti-porcine epidemic diarrhea virus monoclonal antibody, an immunogen and application thereof. The amino acid sequences of complementary determining regions CDR1, CDR2 and CDR3 of a heavy chain variable region of the monoclonal antibody or the antigen binding fragment of the monoclonal antibody are respectively as follows: GFSLSTSGVV, RIDPSDSYT and ARDREKSDNSGYFDY, and the amino acid sequences of complementary determining regions CDR1, CDR2 and CDR3 of a light chain variable region of the monoclonal antibody or the antigen binding fragment of the monoclonal antibody are respectively as follows: RASKSISKYLA, EASSLES and LQHTPKKNK. The monoclonal antibody can be specifically combined with nucleocapsid proteins (N proteins) of all subtypes of the porcine epidemic diarrhea virus, and a porcine epidemic diarrhea virus detection kit prepared from the monoclonal antibody has the advantages of high sensitivity, strong specificity, wide detection range, short detection time and the like.

The invention belongs to the technical field of biological medicine, and discloses a nano antibody for resisting porcine delta coronavirus nucleocapsid protein as well as a recombinant, preparation and application of the nano antibody. According to the nano antibody for resisting the porcine delta coronavirus nucleocapsid protein and the recombinant nano antibody thereof, the specific nano antibody 62Nb targeting the PDCoV N protein is obtained through screening, efficient intracellular delivery of the nano antibody 62Nb is realized through engineering modification of a fusion cell-penetrating peptide TAT, and meanwhile, a bacillus subtilis secretory expression system is established to realize large-scale preparation of 62Nb and a recombinant thereof; finally, the replication process of the PDCoV is inhibited in an intracellular expression or exogenous delivery mode. Meanwhile, the invention provides a large-scale preparation method and application of the nano antibody, and provides a new technical scheme for antiviral prevention and detection of PDCoV.

This invention discloses a kit for detecting porcine reproductive and respiratory syndrome virus (PRRSV) antibodies and its applications, belonging to the field of animal disease detection technology. The kit uses a mutant nucleocapsid protein (N protein) of the PRRSV-2 class NADC30 strain CHsx1401 as the coating antigen; the N protein mutant has mutations at amino acid positions 23, 75, and / or 90 compared to the wild-type N protein. High purity is achieved through optimized prokaryotic expression systems and purification processes. Under optimal detection conditions, the kit exhibits strong specificity, high sensitivity, and good reproducibility, with significantly superior detection performance compared to existing commercial kits. It can be used for PRRSV-2 antibody detection, epidemiological surveys, and disease control.

The invention belongs to the technical field of medicines, and particularly relates to a compound for targeted CSDE1 phase separation and application of the compound in virus resistance. According to the invention, it is found for the first time that the compound 5350-0763 is directly combined with the CSDE1 protein to inhibit the interaction between the compound 5350-0763 and RNA, so that the formation of a CSDE1-RNA aggregate is destroyed. By destroying CSDE1 agglomerates and releasing isolated immune stimulation RNA, RIG-I and other sensors can effectively recognize, so that virus immune escape is relieved, and host innate immune response is restarted. Meanwhile, co-phase separation of CSDE1 and SARS-CoV-2 nucleocapsid protein can be destroyed, and the immune antagonism ability of the virus is weakened. According to the invention, a new medicinal application is developed for the compound 5350-0763, and an experimental foundation is laid and a new view is provided for developing efficient and specific anti-RNA virus medicines, so that the compound 5350-0763 has important clinical significance and social benefits.

The application discloses a kind of laser-induced graphene immune rapid low-cost remote medical detection platform, it is related to biomedical detection technical field, the platform includes following component part: sensor array module: using CO2 laser engraving technology is formed on polyimide substrate and contains 4 graphene working electrode, 1 Ag / AgCl reference electrode and 1 graphene counter electrode patterned structure, through 1-pyrene butyric acid functionalization after specific antibody is fixed and is blocked with bovine serum albumin;The application greatly reduces the detection cost by using laser engraving graphene electrode preparation process, at the same time, by constructing the sensor array module comprising four graphene working electrode, a plurality of antibodies can be specifically fixed, the simultaneous detection of SARS-CoV-2 nucleocapsid protein, spike protein specific IgM / IgG antibody and C-reactive protein and other multiplex biomarkers is realized, sample detection cycle can be controlled within 10 minutes, greatly improve detection efficiency.

A method and composition for the simultaneous detection of a protein and a nucleic acid within a single biological sample using proximity ligation of aptamers and quantitative polymerase chain reaction (qPCR) is disclosed. The invention employs two aptamers, each targeting a distinct epitope of the protein of interest, functionalized with primer sites to enable ligation by way of a short DNA linker in the presence of the protein. This ligated construct is then amplified and detected using qPCR. Concurrently, viral RNA is detected in the same reaction using reverse transcription- qPCR (RT-qPCR) with TaqMan probes in a separate fluorescent channel. This dual detection approach is demonstrated with SARS-CoV-2 nucleocapsid protein and RNA and is validated in saliva samples and digital droplet PCR. The invention provides a sensitive, rapid, and specific method for multiplexed molecular diagnostics.

The invention provides a fusion protein of flounder netshell virus nucleocapsid protein, bacillus subtilis for expressing the fusion protein and an oral vaccine, and relates to the technical field of biology. According to the present invention, the sequence of the bacillus subtilis anchoring protein CotC is obtained through sequencing, the bacillus subtilis anchoring protein CotC and the flounder nested virus are fused, and after the fusion protein is introduced into the bacillus subtilis, the fusion protein can be anchored on the surface of the bacterial spore, and can be used for preparing the recombinant oral vaccine capable of being stably stored and directly fed, and a new method is developed for preventing the flounder netlike virus CSBV.

The invention provides a nucleocapsid protein mutant, a gene, a recombinant expression vector, a mutant virus strain and application, and belongs to the technical field of biological medicine. On the basis of an SARS-CoV-2 virus-like particle (VLP) pseudotyping technology, it is found that N protein alanine mutation occurs, a hydrophobic pocket of N protein CTD is destroyed by L353A, H356A, I357A, D358A and Y360A, and the nucleic acid binding affinity is reduced; it is also found that F403A, F407A and M411A lead to increase of N protein monomers, decrease of tetramers and increase of abnormally high poly N protein, and replication of SARS-CoV-2 wild living viruses and variants is inhibited. Meanwhile, the chiral D-type polypeptide is designed for the CTD region, and it is found that the chiral D-type polypeptide can effectively inhibit assembling of the VLP pseudovirus.

The invention relates to animal virology and genetic engineering technologies, and aims to provide a recombinant porcine epidemic diarrhea virus with enhanced replication capability. The virus is obtained by modifying a porcine epidemic diarrhea virus PEDV, and the amino acid sequence of nucleocapsid protein (N protein) of the virus is shown as SEQ ID NO: 2. The strain of the virus is preserved in the China Center for Type Culture Collection, the preservation name is rcPEDV P120-NT139A, and the preservation number is CCTCC NO: V202562. According to the invention, threonine at the 139th site of nucleocapsid protein of PEDV is a key modification site influencing PEDV infection, and the new strain can be successfully infected in PEDV infection model verification experiments of IPEC-J2 and Vero cells; the strain shows stronger replication ability in a PEDV infection model verification experiment of IPEC-J2 cells, the production difficulty is reduced, and the strain can be further used for preparing a porcine epidemic diarrhea virus inactivated vaccine.

The invention belongs to the technical field of virus immunity, and particularly relates to a hantavirus nucleocapsid protein pan-MHC-I epitope peptide, a vaccine and application. The pan-MHC-I epitope peptide of the hantavirus nucleocapsid protein is at least one of amino acid sequences as shown in SEQ ID NO. 1 to SEQ ID NO. 4. The epitope peptide is used for preparing the vaccine, the immune effect of the vaccine is verified, and meanwhile, the epitope peptide is used for evaluating the effectiveness of the vaccine in the prior art. The invention clarifies the immunological characteristics of hantavirus NP antigens and the application potential of the hantavirus nucleocapsid protein pan-MHC-I epitope peptides in virus infection, disease progression and prevention. The hantavirus nucleocapsid protein pan-MHC-I epitope peptide and the corresponding vaccine are developed by using the MHC-I epitope, and CD8 + T cell response can be remarkably enhanced and activated, so that the defect of singly using the MHC-II epitope is overcome.

The application provides a T cell immune dominant epitope peptide of a SARS-CoV-2 nucleocapsid protein mutation site and an application thereof, and belongs to the technical field of biological medicines. The application develops a dominant epitope peptide as shown in SEQ ID NO. 12 which can specifically target a mutation site and activate a T cell response by screening and identifying a dominant epitope for the N protein of the Omicron mutant strain JN.1 subline. The epitope peptide can induce the body to produce an effective specific T cell immune response against the currently prevailing Omicron subline JN.1 subline mutation site, and enhance the cellular immune capacity of the body. A novel coronavirus vaccine, a drug or an antigen detection kit developed based on the epitope peptide complex can provide a possibility for effectively eliminating virus-infected cells, reducing the persistence of viruses in the body and identifying coronavirus infection, thereby early diagnosing, preventing and treating SARS-CoV-2 breakthrough infection and reducing the long-term health burden of patients.

The invention discloses preparation of an akabane disease virus N protein monoclonal antibody and a double-antibody sandwich ELISA (Enzyme-Linked Immunosorbent Assay) detection method, in order to prepare the akabane disease virus nucleocapsid protein monoclonal antibody, an eukaryotic recombinant plasmid pFastBac-AKAV-N is constructed and is converted into a DH10bac competent cell, and a recombinant stem Bacmid-AKAV-N for expressing AKAV N is successfully obtained. After the recombinant baculovirus is transfected to an sf9 insect cell, the recombinant baculovirus capable of stably expressing the AKAV N protein is rescued. A mouse and a rabbit are immunized with the identified and purified recombinant N protein, and a monoclonal antibody and a polyclonal antibody are prepared respectively. The titer of the polyclonal antibody is measured through an ELISA method, and two positive cell strains are obtained through screening and are named as 2G4 and 6C7 respectively. Furthermore, the prepared polyclonal antibody is used as a coating antigen, an HRP-labeled 2G4 monoclonal antibody is used as a detection antibody, and a double-antibody sandwich ELISA detection method for AKAV N protein antigen detection is optimized and established. According to the method, the AKAV N antigen can be specifically recognized, and reliable technical support is provided for clinical antigen monitoring of AKAV. The monoclonal antibody preparation system and the double-antibody sandwich ELISA detection method established by the invention lay an important foundation for pathogen detection and epidemiological investigation of akabane disease.

The application relates to the technical field of immunological detection, and particularly relates to a mouse anti-swine epidemic diarrhea virus monoclonal antibody, an immunogen thereof and application. The amino acid sequences of the complementarity determining regions CDR1, CDR2 and CDR3 of the heavy chain variable region of the monoclonal antibody or antigen binding fragment thereof are GFSLSTSGVV, RIDPSDSYT and ARDREKSDNSGYFDY respectively, and the amino acid sequences of the complementarity determining regions CDR1, CDR2 and CDR3 of the light chain variable region are RASKSISKYLA, EASSLES and LQHTPKKNK respectively. The monoclonal antibody can specifically bind to the nucleocapsid protein (N protein) of all subtypes of the swine epidemic diarrhea virus, and a swine epidemic diarrhea virus detection kit prepared by using the monoclonal antibody has the advantages of high sensitivity, strong specificity, wide detection range, short detection time and the like.

The application belongs to the technical field of biological detection, and particularly relates to a combination of monoclonal antibodies against nucleocapsid protein of canine distemper virus and application. The combination is composed of monoclonal antibodies 1C8 and 1G8, and the sequences of the heavy chain and light chain variable region complement determining regions are clear (as shown in SEQ ID NO. 1-12, respectively), which endow the combination with high affinity and specificity. It has been verified that the combination can specifically recognize CDV-NP, and has no cross reaction with irrelevant antigens such as canine parainfluenza virus, canine coronavirus and canine parvovirus, thereby guaranteeing the specificity of detection from the source. The antibody combination can be used for preparing colloidal gold detection test strips, kits and other detection tools. The colloidal gold detection test strip based on the double antibody sandwich method uses 1C8 as a capture antibody and 1G8 as a labeled antibody, and is simple to operate, intuitive in results, does not require professional equipment, and is suitable for rapid screening in the field.

Relate to an isolated vNAR single domain antibody that specifically binds to SARS-CoV-2 nucleocapsid protein, and a process for preparing the same.This isolated vNAR single domain antibody are also included.

Provided are compositions and methods for introducing proteins into cells. The compositions and methods relate to introducing a foreign protein as an engineered component of a paramyxovirus virus like particle (VLP). The compositions and methods pertain to modified VLPs that contain a contiguous recombinant polypeptide comprising i) all or a segment of a C-terminal domain of a paramyxovirus nucleocapsid protein and ii) a polypeptide sequence of a distinct protein that is an enzyme such as a recombinase.

This disclosure provides recombinant fusion proteins for use in relation to the novel coronavirus (COVID-19), comprising a SARS-CoV-2 nucleocapsid protein (N-protein) fragment or an analogue linked to a human Fc fragment. Multiple embodiments include administration of the fusion protein as a booster vaccine to patients recovering from COVID-19, administration to antibody-naive patients to produce antibodies against the SARS-CoV-2 virus so that the patient can become a convalescent plasma donor, administration to patients infected with the SARS-CoV-2 virus and suffering from COVID-19 to limit the scope of infection and improve the disease, and administration as a prophylactic COVID-19 vaccine. Exemplary Fc fusion proteins and pharmaceutical formulations of exemplary Fc fusion proteins are provided in addition to methods of use and preparation.

This invention provides an antibody or fragment thereof that specifically binds to the NTD or CTD of the SARS-CoV-2 N protein, and its applications. [Solution] An antibody or antigen-binding fragment thereof that specifically binds to the NTD or CTD of the SARS CoV2 N protein, which consists of a specific amino acid sequence, is provided.

The invention belongs to the technical field of medicine preparation, and particularly discloses an interference peptide for inhibiting mitochondrial autophagy induced by coronavirus nucleocapsid protein as well as a preparation method and application of the interference peptide. The interference peptide (named as ISQOR beta2) is composed of D-type amino acids, and the structure of the interference peptide comprises a polypeptide sequence as shown in SEQ ID NO: 1; the 5-fluorescein isothiocyanate is covalently connected to the N end of the polypeptide. By competitively blocking the interaction between IBV-N and SARS-CoV-2-N and quinone sulfide oxidoreductase, mitochondrial autophagy activation, H2S accumulation in cells and mitochondrial potential depolarization are effectively inhibited, and the expression of innate immune molecules is recovered, so that virus replication is resisted, and lung injury is relieved. The invention solves the problems of difficulty in blocking protein interaction, poor peptide stability and incapability of effectively recovering immune pathways in the prior art, and has the advantages of high in-vitro and in-vivo stability, small toxic and side effects, large patent medicine potential and the like.

The application provides an atomized inhalation new crown vaccine and a preparation method thereof, the new crown vaccine is a recombinant protein, and a nucleotide sequence encoding the recombinant protein is obtained by connecting the following modules in series: a SP signal peptide nucleic acid artificial sequence, a S protein nucleic acid artificial sequence, a Tri T4 phage fibroin folding region nucleic acid artificial sequence, a Linker nucleic acid artificial sequence, a nucleic acid artificial sequence of a Fc antibody constant region, and a N nucleocapsid protein nucleic acid artificial sequence. In the selection process of the immunization antigen, in addition to the S protein, the relatively conservative nucleocapsid protein is also selected as the immunization antigen, so that the effective protection of the vaccine on the body can be maintained in the case of mutation of the new crown virus. The vaccine prepared by the application belongs to an inhalation new crown vaccine, can target the respiratory mucosa immunity through FcRn, and can enhance the immune protection efficiency of the vaccine.

The application belongs to the technical field of medicine preparation, and specifically discloses an interfering peptide for inhibiting mitochondrial autophagy induced by a coronavirus nucleocapsid protein, and a preparation method and application thereof. The interfering peptide (named ISQORbeta2) is composed of D-type amino acids, and the structure comprises: a polypeptide sequence as shown in SEQ ID NO:1; and 5-isothiocyanate fluorescein covalently connected to the N terminal of the polypeptide. The interfering peptide effectively inhibits mitochondrial autophagy activation, intracellular H2S accumulation, mitochondrial potential depolarization, and restores the expression of innate immune molecules by competitively blocking the interaction between IBV-N and SARS-CoV-2-N and sulfide quinone oxidoreductase, thereby resisting viral replication and reducing lung injury. The application solves the problems of the prior art, such as difficulty in blocking protein interaction, poor peptide stability, and inability to effectively restore the immune pathway, and has the advantages of high in-vitro and in-vivo stability, small toxic and side effects, and great potential for drug development.

The application belongs to the technical field of biological detection, and discloses a pig delta coronavirus lateral flow immunochromatographic detection test strip based on nanobodies and application thereof. The test strip takes PDCoV nucleocapsid protein as a detection target, adopts a sandwich type immunodetection principle, and the core recognition element is two high-affinity nanobodies recognizing different epitopes of PDCoV N protein. 62Nb is fused with a pig IgG Fc fragment to form 62Nb-pFc, and colloidal gold is coupled as a detection probe; 94Nb is fixed on the T line as a capture antibody, and the C line is fixed with a goat anti-pig IgG. The nanobodies are prepared by a Pichia pastoris secretion expression system. The test strip is simple to operate and does not require instruments, and can complete visual detection of PDCoV in complex samples such as pig fecal swabs and breeding environment wastewater in 10-15 minutes. The test strip has high sensitivity, strong specificity, good stability, good batch consistency, and is suitable for on-site screening and biological safety monitoring in breeding farms.

The application discloses a kind of monoclonal antibody or its antigen-binding fragment of anti Oya virus nucleocapsid protein and application thereof.The monoclonal antibody F3E10 provided in the application has the characteristics such as high purity, high specificity, high affinity and high sensitivity, can specifically recognize the NP protein of OYAV, can be used as the detection antibody of Western blot and indirect immunofluorescence, is used for the qualitative detection of OYAV-NP protein and OYAV, and can realize the visualization marking and tracing of OYAV under microscope in combination with the secondary antibody labeled with fluorescence, is expected to be widely used in the research, detection and prevention and control of OYAV, and has important popularization and application value in the field of novel bunyavirus research.

Designed coronavirus polypeptide sequences are described, and their use as vaccines against viruses of the coronavirus family. The designed sequences include designed coronavirus spike(S) proteins and fragments thereof, including designed S protein receptor binding domain (RBD) sequence SEQ ID NO:17, designed truncated S protein sequence SEQ ID NO:15, and designed full length S protein sequence SEQ ID NO:13. Designed coronavirus envelope (E), membrane (M), and nucleocapsid (N) protein sequences are also described, and their use as vaccines. Nucleic acid molecules encoding the polypeptides, vectors, fusion proteins, pharmaceutical compositions, cells, and their use as vaccines against viruses of the coronavirus family are also described.

The invention discloses a rabbit-derived single B cell antibody of porcine reproductive and respiratory syndrome virus (PRRSV) nucleocapsid protein, a fusion protein, a detection kit and a detection method. The invention provides two rabbit-derived single B cell antibodies T-11 and T-12, wherein amino acid sequences of variable regions of a heavy chain and a light chain of the rabbit-derived single B cell antibodies T-11 and T-12 are respectively shown as SEQ ID No.1-4; also provided are PRRSV-1 type and type 2 nucleocapsid protein fusion proteins BiN-2 (SEQ ID No.5) and antibody compositions comprising T-11 and T-12. Based on the antibody and the fusion protein, a competitive ELISA detection method and a kit are established, T-11 is used for capturing BiN-2 to construct an indirect coating antigen, biotin-labeled T-12 is used as a competitive antibody, and a judgment result is detected through an enzymatic reaction and a light absorption value. The method can be used for simultaneously detecting the PRRSV-1 type and the PRRSV-2 type antibodies, the sensitivity and the specificity both reach 97% or above, the method has no cross reaction with other porcine virus antibodies, the operation is simple and convenient, the stability is good, and a reliable tool is provided for accurate prevention and control of PRRS and epidemiological monitoring.

This invention relates to the field of biomedical technology, specifically to the application of forsythoside in the preparation of drugs for the prevention and treatment of infectious bronchitis in chickens. Immunofluorescence and Western blotting results show that forsythoside can significantly inhibit the expression of viral nucleocapsid proteins in CEK cells and reduce the viral load in CEK cells. Furthermore, forsythoside can protect against IBV infection in broiler chickens. Quantitative fluorescence results indicate a significant reduction in viral replication in the tracheal tissue of broilers. Clinical symptoms and histopathological results show that forsythoside can significantly alleviate IBV infection-induced slow weight gain, respiratory distress, and tracheal tissue damage in broilers. This invention demonstrates that forsythoside has low cytotoxicity, can effectively prevent infectious bronchitis in chickens, and has advantages such as being green and safe, leaving no residue, having few toxic side effects, and being unlikely to induce drug resistance.

The present disclosure relates to compositions and methods for vaccinating a subject against multiple SARS-CoV-2 variants that involves the making and delivery of extracellular vesicles expressing on their surface engineered spike protein and / or engineered nucleocapsid protein to the subject. The present invention also relates to compositions and methods for the design, preparation, manufacture, formulation, and / or use of spike-display and nucleocapsid-display vesicular vaccines designed to elicit strong humoral and cellular immune responses against multiple SARS-CoV-2 variants.

The invention discloses a small nucleic acid drug delivery system and a preparation method and application thereof. The small nucleic acid drug delivery system includes an adenovirus, a small nucleic acid, and a cleavable linker covalently linking the adenovirus and the small nucleic acid, the cleavable linker including a protease-sensitive functional group or an acidic PH-sensitive functional group. The small nucleic acid drug provided by the invention is connected to sulfydryl of adenovirus nucleocapsid protein through a linker, and the connection mode does not reduce the infection ability of adenovirus to cells. In an in-vitro experiment, the adenovirus can efficiently deliver a small nucleic acid drug, the adenovirus and small nucleic acid can be dissociated after entering a cell, and the small nucleic acid is promoted to enter cytoplasm to play a gene regulation role. In in-vivo and in-vitro experiments, the gene regulation effect of adenovirus delivery is stronger than that of LNP delivery of small nucleic acid drugs, and no toxic effect exists. In a mouse tumor-bearing model, the anti-tumor effect of adenovirus delivery is more remarkable than that of LNP delivery of small nucleic acid drugs.