69 results about "Nucleocapsid Proteins" patented technology

The invention discloses a method for preparing human metapneumovirus nucleocapsid protein based on an insect-baculovirus system. Comprising the following steps: 1) constructing a recombinant vector with a human metapneumovirus nucleocapsid protein coding sequence: sequentially connecting a sequence for promoting secretory expression, the human metapneumovirus nucleocapsid protein coding sequence and a tag sequence from 5'to 3 'ends in the recombinant vector; 2) converting the recombinant vector with the human metapneumovirus nucleocapsid protein coding sequence constructed in the step 1) into a competent cell containing a baculovirus genome plasmid Bacmid to obtain a recombinant baculovirus genome plasmid; and 3) transfecting insect cells with the recombinant baculovirus genome plasmid obtained in the step 2), culturing and purifying to obtain the human metapneumovirus nucleocapsid protein. The invention provides the preparation method of the human metapneumovirus nucleocapsid protein, and a material support is provided for subsequent further research.

The invention relates to the technical field of biological genetic engineering, in particular to construction of recombinant lactobacillus plantarum of an African swine fever virus antigen, and the method comprises the following steps: constructing a lactobacillus plantarum shuttle vector containing different ASFV antigen genes; and referring to a genome sequence of a Chinese isolate CADCHN09 of ASFV disclosed in GenBank. The method has the advantages that an inducer is not needed, antigen protein is efficiently expressed, the dendritic cell (DC) binding peptide for enhancing immune cell uptake and processing is subjected to fusion expression, the oral immunization mode is simple, mucosal immunity is stimulated, and ASFV infection is effectively prevented and controlled; probiotics, namely lactobacillus plantarum, which naturally exist in animal intestinal tracts and can be colonized and copied in digestive tracts for a long time are used as a carrier; key epitopes of proteins such as nucleocapsid protein p72, outer envelope protein CD2v, inner envelope protein p54, p22, pE199L and pE248R, which play an important role in adsorption, internalization, endocytosis and membrane fusion of ASFV, are preferably fused with dendritic cell binding peptide and then are displayed on the surface of lactobacillus plantarum.

The invention relates to a porcine reproductive and respiratory syndrome virus (PRRSV) dimer nucleocapsid (N) protein (PRRSV N-dimer) and application thereof. The PRRSV N-dimer is formed by connecting N protein amino acid sequences in series through (GGGGS) 3linker. According to the invention, a GGGGS 3linker nucleotide sequence is connected in series with a PRRSV NADC30-like strain N protein gene sequence to construct a prokaryotic expression vector, and a recombinant PRRSV N-dimer is expressed in escherichia coli. Through SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis) and Western-blot detection, the recombinant PRRSV N-dimer obtains soluble expression and can be subjected to a specific reaction with PRRSV positive serum; through detection by a freeze thawing method and an ELISA (Enzyme-Linked Immunosorbent Assay) method, compared with a PRRSV monomer N protein (PRRSV N-monomer), the PRRSV N-dimer protein can be stably preserved, and the immunoreactivity is better. A PRRSV antibody indirect enzyme-linked immunosorbent assay (iELISA) detection method is established on the basis of a PRRSV N-dimer, and can be applied to diagnosis and monitoring of PRRSV infection.

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 present disclosure relates to compositions and methods for vaccinating a subject against a variety of SARS-CoV-2 variants and other respiratory viruses, the compositions and methods involve preparing and delivering extracellular vesicles expressing on its surface an engineered spike protein, an engineered nucleocapsid protein, an engineered hemagglutinin protein, and / or an engineered respiratory syncytial virus pre-fusion or fusion (RSV F) protein. The invention also relates to compositions and methods for designing, preparing, making, formulating and / or using spike display, nucleocapsid display, hemagglutinin display and / or RSV F display vesicle vaccines, the spike display, nucleocapsid display, hemagglutinin display, and / or RSV F display vesicle vaccines are designed to elicit potent humoral and cellular immune responses against a variety of respiratory viruses and variants.

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 present invention relates to a prime editing system with enhanced correction precision and a use thereof. More specifically, the present invention relates to a recombinant vector for prime editing with improved precision, comprising: a prime editor expression cassette including a mutant Cas9 (R221K, N394K, D840A), a nucleocapsid protein, and an RNaseH domain-deleted mutant MMLV reverse transcriptase (D200N, T306K, W313F, T330P, L603W); and a pegRNA (prime editing guide RNA) expression cassette transcriptionally regulated by a U6 composite promoter.

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.

Disclosed herein are methods for inducing immunity against a virus such as a coronavirus in the mucosal tissue of a patient, include administering a vaccine composition to the patient by oral administration (e.g., nasal injection, nasal inhalation, oral inhalation, and / or oral ingestion). Also disclosed are compositions for assaying the presence of anti-viral antibodies induced by the administered vaccine or the presence of viral proteins in a saliva sample include a stabilizing solution and may also include the use of aragonite particle beads. Compositions and methods are presented for prevention and / or treatment of a coronavirus disease wherein the composition comprises a recombinant entity. The recombinant entity is bivalent, comprising a nucleic acid encoding a coronavirus 2 nucleocapsid protein CoV2 nucleocapsid protein fused to an endosomal targeting sequence, and a nucleic acid encoding a CoV2 spike protein sequence optimized for cell surface expression.

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 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.

Compositions and methods are presented for prevention and / or treatment of a coronavirus disease wherein the composition comprises comprises a recombinant entity. The recombinant entity comprises a nucleic acid that encodes a nucleocapsid protein of coronavirus 2 (CoV2); and / or wherein the recombinant entity encodes a spike protein of CoV2.

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.

The invention discloses a monoclonal antibody for resisting novel bunyavirus nucleocapsid protein (NP). The monoclonal antibody comprises a heavy chain variable region and a light chain variable region, the amino acid sequence of the heavy chain variable region is as shown in NO.1; the coding DNA sequence of the heavy chain variable region is as shown in NO.2; the amino acid sequence of the light chain variable region is as shown in NO.3; and the coding DNA sequence of the light chain variable region is as shown in NO.4. The monoclonal antibody for resisting the novel bunyavirus NP protein can be specifically combined with the novel bunyavirus NP protein, and a basis is established for diagnosing virus infectious diseases through a genetic engineering method; the monoclonal antibody for resisting the novel bunyavirus NP protein has a specific antigen binding structural domain, can be combined with the novel bunyavirus NP protein in a targeted manner, and is used for antigen detection.

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 invention relates to a bioengineering technology, and aims to provide an IPEC-J2 cell strain for stably expressing a PED virus nucleocapsid protein truncated fragment and application of the IPEC-J2 cell strain. The cell strain is preserved in the China Center for Type Culture Collection, the preservation name is a porcine small intestine epithelial cell line R58-WT-IPEC, the Latin name is Sus scrofa, and the preservation number is CCTCC NO: C202425. The invention discloses a key effect of the PEDV N protein in a virus replication stage, and provides application of the PEDV N protein as a target spot in regulation and control of PEDV infection. Compared with a wild-type IPEC-J2 cell line, the cell line provided by the invention has a remarkable promotion effect, can solve the problem of poor proliferation of PEDV (porcine epidemic diarrhea virus) in the IPEC-J2 cell line, and further improves the application possibility of the IPEC-J2 cell line in virus separation and propagation.