47 results about "Nucleocapsid Proteins" patented technology

The present invention relates to treatment of COVID-19 and methods thereof. A vaccine composition for inducing immunity against coronavirus in a subject comprises a recombinant nucleic acid encoding an N-ETSD, which is a modified nucleocapsid protein comprising an endosomal targeting sequence, and / or a recombinant nucleic acid encoding an S-fusion, which is a modified spike protein with improved surface expression. The vaccines can be formulated as recombinant nucleic acids, recombinant yeasts, and / or recombinant viruses, such as adenoviruses, and can be administered via injection and / or mucosal delivery.

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 present application discloses the use of TIAR inhibitors for treating hepatitis B virus infection. The present application demonstrates that the host factor TIAR promotes HBV replication by regulating the translation of pgRNA through its binding, i.e. inhibiting the translation of nucleocapsid protein but promoting the translation of Pol. It is also found that TIAR is also packaged into the nucleocapsid of HBV through its binding to pgRNA. The present application provides a new solution for treating HBV infection based on TIAR as a target by reducing the expression of TIAR or inhibiting the binding of TIAR to pgRNA to inhibit HBV replication.

A homogeneous method for detecting the nucleocapsid protein of the new coronavirus based on a nano-antibody-monoclonal antibody sandwich relates to the field of antigen detection. The present invention prepares a variety of novel coronavirus nucleocapsid protein nano-antibodies as solid-phase antibodies, nucleocapsid protein monoclonal antibodies as detection antibodies, and novel coronavirus N protein as detection targets. A group of antibody pairs with excellent detection performance are screened out by enzyme-linked immunosorbent assay, with a minimum detection line of 0.8 ng / mL. In addition, a one-step activation method couples nano-antibodies and monoclonal antibodies to the surfaces of quantum dot microspheres and magnetic nano-microspheres, respectively, and successfully prepares two probes targeting the nucleocapsid protein of the new coronavirus. It also constructs a magnetic separation immunohomogeneous method for rapid detection of the nucleocapsid protein of the new coronavirus, with a minimum detection line of 0.15 ng / mL. The present invention is simple to operate, low in cost, fast in reaction speed, can realize in situ detection, and has high efficiency in paired antibody screening. It has good application prospects in the rapid detection of novel coronaviruses.

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 present invention discloses a highly sensitive detection kit for the nucleocapsid protein of the novel coronavirus. The detection kit comprises the capture antibody 6C3-D8 and the detection antibody 5F2-C11, which specifically target the novel coronavirus N protein. The present invention provides a novel method for detecting the nucleocapsid protein of the novel coronavirus in the art, which has high specificity and sensitivity and has broad application prospects.

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 provides a recombinant LLC-PK1 cell line capable of stably expressing a TGEV-N gene and application of the recombinant LLC-PK1 cell line in promotion of virus rescue, and belongs to the technical field of gene engineering. The invention provides a recombinant LLC-PK1 cell line capable of stably expressing TGEV-N. The recombinant LLC-PK1 cell line capable of stably expressing TGEV-N. The recombinant LLC-PK1 cell line capable of stably expressing TGEV-Nis obtained by cloning a TGEV-N fragment into a lentiviral vector to construct a recombinant lentiviral plasmid and infecting LLC-PK1 cells through lentivirus packaged by virus rescue. The recombinant LLC-PK1 cell capable of stably expressing the TGEV-N is beneficial to rescue of TGEV infectious cloned viruses, so that a tool is provided for researching a virus pathogenic mechanism and a novel genetic engineering vaccine.

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.

[Problem] To provide an antiviral oligonucleotide that acts on cells infected by viruses having multiple RNA genomes, exhibiting an antiviral effect, and does not tend to the emergence of resistant viruses, and a pharmacological composition thereof. [Solution] An oligonucleotide that takes on a G-quadruplex structure that inhibits the binding of the genomic RNA and nucleocapsid (N) protein of a virus having an RNA genome, the antiviral oligonucleotide suppressing proliferation of the virus by contact with virus-infected cells, and a pharmacological composition thereof are used.

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 provides a T cell immunodominance epitope peptide of an SARS-CoV-2 nucleocapsid protein mutation site and application of the T cell immunodominance epitope peptide, and belongs to the technical field of biological medicine. According to the invention, a group of dominant epitope peptide compounds capable of specifically targeting mutation sites and activating T cell response are developed by screening and identifying dominant epitopes aiming at N protein of an Omicron mutant strain JN.1 substrain. The epitope peptide can induce an organism to generate an effective specific T cell immune response aiming at a current main epidemic Omicron pedigree JN.1 substrain mutation site, so that the cellular immunocompetence of the organism is enhanced. A novel coronavirus vaccine, a medicine or an antigen detection kit developed based on the epitope peptide compound can provide possibility for effectively removing virus infected cells, reducing continuous existence of viruses in vivo and identifying new coronavirus infection, so that SARS-CoV-2 breakthrough infection is diagnosed and prevented in an early stage, and the long-term health burden of a patient is relieved.

Disclosed are a preparation method and application of a single-chain antibody fragment targeting SARS-CoV-2 nucleocapsid protein, aiming to provide a single-chain antibody fragment with high affinity and strong specificity. The present disclosure obtains three single-chain antibody fragments targeting SARS-CoV-2 nucleocapsid protein through construction of a nanobody phage library and phage screening technology. The single-chain antibody fragment consists of a heavy chain variable region, a 15aa connecting peptide, and a light chain variable region connected in sequence, and has an amino acid sequence shown in SEQ ID NO: 1-SEQ ID NO: 3. The single-chain antibody fragment in the present disclosure has excellent binding performance and specificity with the SARS-CoV-2 nucleocapsid protein, can be applied to a variety of immunoassay platforms of the SARS-CoV-2, and has broad application prospects in the field of SARS-CoV-2 detection.

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.

A method for preparing protein-based nanoparticle for self-packaging and delivering mRNA includes the following steps. A first donor plasmid, a second donor plasmid and a third donor plasmid are provided. A plasmid transposing step is performed. A recombinant virus preparing step is performed so as to obtain a first recombinant baculovirus, a second recombinant baculovirus and a third recombinant baculovirus. A transducing step is performed, wherein the first recombinant baculovirus, the second recombinant baculovirus and the third recombinant baculovirus are used to infect a producer cell so as to express a nucleocapsid protein, an envelope protein, an engineered envelope protein and a target RNA, and the nucleocapsid protein, the envelope protein, the engineered envelope protein and the target RNA are self-assembled to form a protein-based nanoparticle for self-packaging and delivering mRNA.

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.