251 results about "F protein" patented technology

The invention discloses a modified bordetella pertussis strain and an application thereof. The modified Bordetella pertussis strain is characterized in that in the Bordetella pertussis strain BAA-589, the BP2992 protein is not expressed, or the inactive BP2992 protein is expressed, the pldA protein is not expressed, or the inactive pldA protein is expressed, and the mla F protein is not expressed, or the inactive mla F protein is expressed. When the Bordetella pertussis strain is treated by combining an ultrasonic method, compared with wild Bordetella pertussis BAA-589, the Bordetella pertussis strain has a remarkable high-foaming phenotype, the OMV secretion amount is remarkably increased, and a good technical foundation is laid for developing a Bordetella pertussis OMV vaccine.

The invention relates to the technical field of biology, in particular to a multivalent pneumococcal polysaccharide-protein conjugate vaccine for preventing pneumococcal infection and respiratory syncytial virus infection as well as a preparation method and application of the multivalent pneumococcal polysaccharide-protein conjugate vaccine. According to the multivalent pneumococcal polysaccharide-protein conjugate vaccine disclosed by the invention, carrier protein is RSV recombinant protein with immunogenicity; the pneumococcal polysaccharide is connected with the RSV recombinant protein through a covalent bond; the RSV recombinant protein is selected from Pre-F protein, and the amino acid sequence of the RSV recombinant protein is shown as SEQ ID NO: 2 or SEQ ID NO: 3. The conjugate vaccine provided by the invention is an innovative vaccine, two common diseases, namely pneumonia and respiratory syncytial virus infection, can be simultaneously prevented by using one vaccine product, and the number of vaccination times of the vaccine is greatly reduced. According to the invention, the carrier protein in the conjugate vaccine is subjected to mutation modification, so that the carrier protein can maintain structural stability and antigen cluster function in different environments.

The invention relates to the field of prevention and treatment of respiratory diseases, and discloses a preparation method of a human respiratory syncytial virus preventive mRNA vaccine. The main components of the mRNA vaccine comprise mRNA for coding an antigen and lipid nanoparticles. According to the mRNA sequence provided by the invention, an independently researched and developed 5 'UTR / 3' UTR sequence and a codon optimization mode are adopted, and efficient translation of target protein can be realized. According to the mRNA vaccine provided by the invention, RSV F protein is selected as an antigen, and the RSV F protein sequence is optimally designed, so that stable pre-fusion conformation RSV F protein can be expressed, and after a mouse is immunized, the mouse can be induced to generate high-level RSV F protein specific binding antibody titer and neutralizing antibody titer.

The invention relates to an anti-RSV polypeptide membrane fusion inhibitor as well as a preparation method and application thereof. The invention discloses a compound, a pharmaceutical salt or a derivative thereof, a pharmaceutical composition and a preparation product containing the compound, and application of the compound and the pharmaceutical salt or the derivative in preparation of medicines for preventing and / or treating diseases caused by RSV or other paramyxoviridae viruses. The polypeptide disclosed by the invention shows the anti-RSV activity and safety which are obviously superior to those of positive control polypeptide. Molecular interaction experiments show that the polypeptide acts on the NHR region of the RSV F protein, and the fusion of the RSV virus and a target cell is prevented by forming an inactive six-helix structure with NHR, or by destroying the RSV NHR structure and inhibiting the process of forming a six-helix bundle by NHR and CHR in the F protein in the key step of RSV infection.

The present invention is generally related to modified or mutated respiratory syncytial virus fusion (F) proteins and methods for making and using them, including immunogenic compositions such as vaccines for the treatment and / or prevention of RSV infection.

The invention relates to the technical field of virology, immunology and biomedicine, in particular to a human metapneumovirus F protein mutant and application thereof. The human metapneumovirus F protein mutant provided by the invention has stable prefusion conformation and high expression quantity, and can form a stable trimer, so that the human metapneumovirus F protein mutant can be used for HMPV vaccine development. The F protein mutant provided by the invention has excellent immunogenicity, and can induce excellent immune response when being used together with an HA201 composite adjuvant (QS21 + MPL liposome).

The invention provides a bovine parainfluenza virus type 3 QL3 strain, and the preservation number of the bovine parainfluenza virus type 3 QL3 strain is CGMCC No.46469. The invention also provides a bovine parainfluenza inactivated vaccine containing the bovine parainfluenza virus type 3 QL3 strain and purified F protein and HN protein of the QL3 strain. The novel bovine parainfluenza virus type 3 inactivated vaccine is obtained by optimizing the antigen components of the bovine parainfluenza virus type 3 inactivated vaccine, and high-level neutralizing antibodies can be generated after immunizing cattle; the safety of the vaccine is improved through process optimization. Safety and potency test results show that after the inactivated vaccine is used for immunizing cattle, no adverse reaction exists, and high-level neutralizing antibodies are generated. Results show that the immune effect of the bovine parainfluenza virus type 3 inactivated vaccine can be improved.

The invention discloses a specific monoclonal antibody for resisting a Nipah virus Post-F protein, the monoclonal antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3; wherein the HCDR1, the HCDR2 and the HCDR3 respectively comprise the HCDR1, the HCDR2 and the HCDR3 in a heavy chain variable region of which the amino acid sequence is any one of SEQ ID NO: 4, 13, 20, 25, 30, 34 or 42, or respectively have at least 85% sequence identity with the HCDR1, the HCDR2 and the HCDR3; and / or the LCDR1, the LCDR2 and the LCDR3 respectively comprise LCDR1, LCDR2 and LCDR3 in a light chain variable region with the amino acid sequence of any one of SEQ ID NO: 9, 16, 23, 28, 32, 37 or 46, or respectively have at least 85% sequence identity with the LCDR1, the LCDR2 and the LCDR3. The anti-Nipah virus Post-F protein monoclonal antibody disclosed by the invention can be specifically combined with a Post-F conformation protein, and has the application potential of being developed into a Nipah virus antibody detection kit or a virus neutralizing antibody drug.

The present invention provides engineered soluble F proteins of paramyxoviruses, such as respiratory syncytial virus (RSV), human metapneumovirus (hMPV), and human parainfluenza virus (hPIV). These engineered proteins are stabilized by specific modifications in the wild-type soluble F sequence, such as substitutions in the 023 chain and / or the introduction of engineered disulfide bonds in the 0 hairpin in the F1 subunit. The present invention also provides a nanoparticle vaccine comprising an engineered soluble F immunogen displayed on a self-assembled nanoparticle. The invention also provides methods of using such vaccine compositions in a variety of therapeutic applications, e.g., methods for preventing or treating viral infections, such as RSV, MPV, and PIV infections.

The invention discloses a reconstructed respiratory syncytial virus F protein trimer, relates to the technical field of genetic engineering vaccines, and designs an RSV F protein trimer, a monomer of the trimer does not contain a p27 sequence, and a furin enzyme digestion peptide fragment aa110-aa136 is deleted; the F2 structural domain and the F1 structural domain are connected through a GS connexon; the invention also designs point mutation combinations, and the point mutation combinations can form an inter-chain disulfide bond, so that covalent connection of the trimer is realized, and the problem of depolymerization of the trimer is fundamentally solved; the trimer is stabilized in a pre-F state, a post-F sample structure does not exist, and an ELISA result shows that a neutralizing epitope is completely reserved; the trimer antigen structure is challenged by adopting a thermal acceleration experiment method, and a result shows that the purity of the trimer antigen reaches 95% or above after the trimer antigen is placed at 37 DEG C for 28 days, and the trimer antigen is very good in stability; when the trimer is used for preparing the vaccine, the standard required by laws and regulations can be met without freeze-drying.

The present invention relates to the technical field of biomedicine. Specifically provided are a respiratory syncytial virus (RSV) Pre-F recombinant protein nanoparticle vaccine and a preparation method therefor. By performing amino acid mutation on an RSV Pre-F protein, and performing fusion expression on the mutated Pre-F protein and ferritin particles in eukaryotic cells, PreF-Ferritin fusion protein nanoparticles, in which eight Pre-F protein trimers are densely displayed on the surface of each ferritin particle, are obtained. By stabilizing and exposing antigen epitopes required to be displayed and disrupting or masking unrequired antigen epitopes, the PreF-Ferritin fusion protein nanoparticles effectively improve the immunogenicity and the production stability of the antigen. Experiments show that: when the PreF-Ferritin fusion protein is injected into mice, high-titer protective sera can be acquired, and the mice sera can yield relatively high neutralizing titers against RSV.

The present invention relates to RSV F protein mutants and their applications, belonging to the field of vaccines. Based on the amino acid sequence of wild-type RSV F protein, the present invention has carried out modifications in multiple aspects and in various ways, including disulfide bond mutations, proline mutations, cavity filling mutations, electrostatic mutations, p27 sequence modifications, and / or trimerization domain modifications, and designed a variety of RSV F protein mutants. Then, through gene synthesis, expression, and ELISA detection, the present invention tested the pre-fusion conformation (pre-F) ratio and stability under high and low temperature conditions of the RSV F protein mutants, and screened out several RSV F protein mutants with a relatively high pre-fusion conformation ratio and stable structure. Finally, candidate antigen representatives were selected for immunogenicity detection, which proved that they have good immunogenicity and can be used for the development of RSV vaccines, the preparation of RSV-specific antibodies, or RSV detection reagents.

The invention discloses an RSV pre-F tripolymer protein combination and application thereof in preparation of bivalent RSV vaccines, and relates to the technical field of human vaccines. The vaccine is prepared by mixing an RSV-A subtype pre-F tripolymer protein and an RSV-B subtype pre-F tripolymer protein in equal mass, and matching with an aluminum hydroxide adjuvant with the concentration of 0.35 mg / 0.5 mL. A p27 fragment and a C-terminal esterification region of each of the two F proteins are deleted, the two F proteins are connected with F2 / F1 through GSGSGS, inter-chain disulfide bonds are introduced to S146C / N460C and A149C / Y458C sites respectively, intra-chain disulfide bonds are introduced to S155C / S290C sites respectively, and stable pre-F trimers are spontaneously assembled after secretion expression of CHO cells; the vaccine does not need to be freeze-dried, still keeps high purity and neutralizing epitope integrity after being stored at 37 DEG C for 28 days, can obviously induce neutralizing antibody and T cell immunity, and can be used for preventing various RSV-A / B infections.

Related to the field of prevention and treatment of respiratory diseases, and disclosed a method for preparing a preventive mRNA vaccine against human respiratory syncytial virus. In some embodiments, the mRNA vaccine comprises an antigen-encoding mRNA and lipid nanoparticles. In some embodiments, the mRNA sequence utilizes an improved 5'UTR / 3'UTR sequence and codon optimization methods for efficient translation of the target protein. In some embodiments, the mRNA vaccine expresses an RSV F protein that is used as an antigen, wherein the sequence of the RSV F protein is optimized to achieve stable RSV pre-F protein expression. After immunization, the mRNA vaccine induces high levels of RSV F protein-specific binding antibody titer and neutralizing antibody titer in mice.

The invention relates to an RSV F protein soluble fragment, an autotrimer structural domain and application thereof, and belongs to the technical field of recombinant protein. Based on an amino acid sequence of a wild RSV F protein, a soluble and rigid sequence is screened out, a soluble fragment is constructed, disulfide bond mutation pair modification is performed on the basis of the soluble fragment to obtain a self-derived trimer structural domain sequence, and the self-derived trimer structural domain sequence is used for replacing a transmembrane region and an intracellular region of the RSV F protein. Therefore, trimerization can be realized, stable pre-fusion conformation can be maintained, and soluble expression in cells can be realized at the same time. The invention also provides an RSV F protein mutant containing an autotrimer structural domain sequence, the generation and the stability of the pre-F conformation trimer are proved, a representative is selected from the pre-F conformation trimer for animal experiments, and it is proved that the pre-F conformation trimer has good immunogenicity and can be used for developing RSV vaccines and preparing RSV specific antibodies or RSV detection reagents.

The invention belongs to the technical field of medicine and biology, and particularly relates to a vaccine for preventing and / or treating respiratory syncytial virus. Respiratory syncytial virus Pre F protein is selected as a skeleton, and the C end of the skeleton is connected with a G protein CCD part fragment. According to the invention, double neutralizing antibodies of RSV preF protein and G protein can be generated at the same time, and double effects of preventing viruses from infecting a host and preventing residual viruses from spreading again are achieved. Further, different adjuvants are utilized to improve humoral immune response and reduce vaccine enhanced pathological injury (VED) at the same time. Therefore, the invention has practical value in preventing or treating respiratory syncytial disease virus infection.

The invention relates to a monoclonal antibody targeting RSV F protein, and a related product and application thereof. The monoclonal antibody (AD5) or the antigen binding fragment of the monoclonal antibody is specifically bound with the Site III antigen epitope of the RSV F protein. The antibody AD5 disclosed by the invention has the characteristics of targeting high-conservative epitope (Site III) and having high neutralizing activity, and overcomes the main defect that the neutralizing activity is not high when the existing listed antibody targets the high-conservative epitope.

The present invention discloses mutants of wild-type RSV F protein. The binding experiment of pre-fusion conformation trimer-specific monoclonal antibody AM14 shows that the mutant provided in the embodiment of the present disclosure can have strong binding activity with AM14 monoclonal antibody, while the wild-type RSV F protein before artificial mutation has no binding activity with AM14 monoclonal antibody. The results of accelerated stability test show that the mutant provided in the embodiment of the present disclosure is placed in a 37°C environment for 4 weeks, and the binding activity of AM14 monoclonal antibody has no obvious change. The results of animal immunization experiments show that compared with the wild-type RSV F protein before artificial mutation, the mutant provided in the embodiment of the present disclosure can induce the production of higher titers of neutralizing antibodies.

The invention relates to the technical field of biological medicine, and particularly discloses a monoclonal neutralizing antibody for resisting respiratory syncytial virus and application of the monoclonal neutralizing antibody. The antibody provided by the invention can identify non-denatured RSV (L) protein through ELISA (enzyme-linked immunosorbent assay), but cannot identify denatured RSV (D) protein, and is a kappa IgG2b type antibody. The molecular marker is 0.0001 mu g / ml-1 mu g / ml, can positively recognize the RSV Pre-F protein, and has a gradient effect. The antibody can be used as a neutralizing active antibody in an ELISA double-antibody sandwich method, can be used for detecting the in-vitro potency of the RSV PreF protein, and plays an important role in evaluating the vaccine activity in the vaccine research and development process.

The invention belongs to the technical field of medicines, and discloses application of an adeno-associated virus vector in preparation of medicines for preventing and treating RSV infection and / or diseases caused by RSV. According to the invention, a recombinant adeno-associated virus vector is adopted for the first time to construct a medicine for preventing and treating RSV infection or diseases caused by RSV infection, and based on this, the invention provides a nucleic acid molecule which comprises: (a1) a first region which comprises a first adeno-associated virus inverted terminal repeat sequence, and (a2) a second region which comprises a second adeno-associated virus inverted terminal repeat sequence; and (a2) a second region comprising a gene encoding a respiratory syncytial virus antigen; the recombinant adeno-associated virus containing the nucleic acid molecule can generate better humoral immune effect and cellular immune effect, has a certain serum neutralization effect on strains of A and B subtypes of RSV, and particularly, when the antigen is a fusion protein of the respiratory syncytial virus or a fusion protein mutant of the respiratory syncytial virus, the recombinant adeno-associated virus has a certain serum neutralization effect on the strains of A and B subtypes of the RSV. The recombinant adenovirus vector containing the nucleic acid molecule can successfully realize the expression of the F protein before RSV fusion.

The invention provides a human metapneumovirus monoclonal antibody and application thereof, and belongs to the technical field of immunity. The monoclonal antibody F720-1 provided by the invention only has binding activity to conformation F protein before human metapneumovirus fusion, and the monoclonal antibody F390-6 has binding activity to conformation F protein before human metapneumovirus fusion and conformation F protein after human metapneumovirus fusion. Through pairing, it is determined that F720-1 and F390-6 can conduct proportion detection on Pre-F and Post-F in the human metapneumovirus F protein through an enzyme-linked immunosorbent assay double-antibody sandwich method and conduct quantification on Pre-F. The double-antibody sandwich ELISA method has a good linear relation and detection limit when used for detecting the proportion of the Pre-F protein to the Post-F protein in the F protein. The method is expected to be used for accurate evaluation in human metapneumovirus protein experimental vaccine stock solution and vaccine verification.

The present invention relates to a respiratory syncytial virus subgroup B (RSV-B) recombinant F protein, a polynucleotide encoding the same, a nucleic acid construct comprising the polynucleotide, an expression vector comprising the nucleic acid construct, a host cell transformed or transfected with the above-mentioned polynucleotide, nucleic acid construct or expression vector, a stabilized trimer formed therefrom, an immunogenic composition comprising any one of the foregoing, and its use in the preparation of a vaccine for preventing and / or treating respiratory syncytial virus infection. The RSV-B recombinant F protein of the present invention comprises at least one specific epitope of the prefusion F protein, can form a stable F protein trimer in the prefusion conformation, and has stable expression, uniform form and greatly improved yield; the formed F protein trimer has good immunogenicity, can stimulate the body to produce a high level of antibody titer, and has great significance for the clinical treatment and prevention and control of respiratory syncytial virus.

The invention discloses a nano antibody of pre-F protein of a targeted respiratory syncytial virus and application of the nano antibody. The nano antibody comprises a heavy chain variable region, and the heavy chain variable region comprises heavy chain complementary determining regions CDR1, CDR2 and CDR3. The nano antibody disclosed by the invention can be specifically combined with RSV, and has a good application prospect in the aspect of RSV infection related diseases.

The invention discloses a respiratory syncytial virus F protein mutant and a vaccine, and belongs to the technical field of biological medicines. The mutant is capable of inducing a higher neutralizing antibody titer against the respiratory syncytial virus. According to the invention, the amino acid sequence of the RSV pre-fusion protein is mutated and transformed, so that the stability is enhanced, the immunogenicity of the vaccine to different RSV variants is improved, and the protection effect of the vaccine is improved.

The invention relates to the technical field of vaccines, and particularly discloses a recombinant extracellular domain mutant trimer of a respiratory syncytial virus pre-fusion F protein, application of the recombinant extracellular domain mutant trimer and a vaccine. The mutant trimer comprises three recombinant F2-F1 extracellular domain protomers, and the recombinant F2-F1 extracellular domain protomers comprise three recombinant F2-F1 extracellular domain In some embodiments, the recombinant F2-F1 extracellular domain protomers each comprise a deletion of positions 104-144 and a peptide linker between positions 103 and 145, and comprise a L373R substitution and an engineered disulfide bond consisting of 55C and 188C, 149C and 458C substitutions, relative to a reference sequence. The recombinant extracellular domain mutant trimer of the F protein before fusion of the respiratory syncytial virus, provided by the invention, has higher yield, and the immunogenicity and stability of the antigen are further improved.

Provided are virus-like particle vaccines for human metapneumovirus (hMPV) in which the ectodomain of hMPV F protein is linked to, and thereby displayed on, a symmetric protein-based virus-like particle. For example, the vaccine antigen may be a N-terminal fusion of the ectodomain of hMPV F protein to a protein having a multimerization domain for a one- or two-component virus-like particle, such as a two-component icosahedral virus-like particle. Further provided are vaccine compositions, methods of manufacturing, and methods of use, e.g., immunizing a subject to generate a protective immune response to hMPV.

The invention discloses a Nipah virus F protein monoclonal antibody and application thereof, the antibody comprises an antibody 2D12 and an antibody 3B10, and the Nipah virus F protein monoclonal antibody is prepared from Nipah virus F protein immunogen. The specific anti-Nipah virus F protein monoclonal antibody pair is prepared and detected, the monoclonal antibody pair can be specifically combined with the Nipah virus F protein, is high in titer and good in affinity and has neutralizing activity, and meanwhile, the detection kit prepared on the basis of the antibody pair can effectively carry out Nipah virus detection; and technical support can be provided for treatment, epidemiological monitoring and diagnosis of Nipah virus infection.

The present invention relates to the field of immunology and molecular virology, and in particular relates to the field of the prevention and treatment of RSV. Specifically, provided are a monoclonal antibody capable of specifically binding to a new epitope between an Ø epitope and a V epitope on a pre-F protein or a new epitope between an II epitope and a V epitope, and the use thereof in the detection, prevention and / or treatment of RSV infection and / or diseases caused by the infections.

The application belongs to the technical field of biological medicine, and particularly relates to a RSV pre-fusion F protein mutant and application thereof. The RSV pre-fusion F protein mutant has good stability, high expression and / or immunogenicity, and can be used for preparing a RSV antibody, preventing RSV infection, preventing and / or treating a disease caused by RSV infection, detecting the presence or content of the RSV antibody in a sample, diagnosing RSV infection, and diagnosing a disease caused by RSV infection.

The invention provides a monoclonal antibody aiming at human metapneumovirus (hMPV) or an antigen binding fragment thereof, a related product thereof, and a preparation method and application thereof. The monoclonal antibody or the antigen binding fragment of the monoclonal antibody can be specifically bound with each subtype F protein of the hMPV with higher affinity, and has higher neutralizing activity on each subtype virus of the hMPV, so that the monoclonal antibody or the antigen binding fragment of the monoclonal antibody has extremely high potential to be developed into an hMPV antigen detection kit and an hMPV broad-spectrum therapeutic antibody; the method shows a remarkable application prospect in the field of clinical detection and treatment of the hMPV, and has an extremely high application value.