567 results about "Neutralising antibody" patented technology

The invention discloses a self-replicating RNA (Ribonucleic Acid) molecule, a porcine rotavirus RNA vaccine and application thereof, and relates to the technical field of vaccines. According to the present invention, the coding sequence of the porcine rotavirus G9P23 type VP4 protein is optimized, such that the fact that the saRNA prepared by using the nucleotide sequence represented by the SEQ ID NO: 10 can secrete the high porcine rotavirus G9P23 type VP4 protein antigen after the cell transfection is found; after the optimized porcine rotavirus G9P23 type VP4 protein is used for immunizing experimental animals, the level of neutralizing antibodies generated by the animals can be improved, and the challenge protection effect is enhanced. Therefore, the nucleic acid molecule provided by the invention can be used for developing drugs and vaccines for causing protective immune response in experimental animal bodies. Compared with a traditional vaccine, the vaccine provided by the invention has good immunogenicity and safety, and has a good application prospect.

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 anti-drug neutralizing antibody analysis. In particular, the present application relates to methods for detecting neutralizing antibodies against a vascular endothelial growth factor (VEGF) binding moiety and a programmed cell death ligand-1 (PD-L1) binding moiety in a biological sample from an individual administered a bispecific antibody drug comprising the VEGF binding moiety.

The present invention belongs to the field of virus detection technology, and specifically relates to monoclonal antibodies with neutralizing activity against adenovirus type 5 and their uses. The monoclonal antibody 8B9 of the present invention can specifically recognize and neutralize adenovirus type 5, and the amino acid sequences of the heavy and light chain variable regions and their complementarity determining regions of the monoclonal antibody 8B9 are provided. The present invention also provides an in vitro neutralization activity assessment model based on HEK-293A cells. At a concentration as low as 3.3 μg / mL, the titer of the neutralizing activity against the virus stock solution after 6000-fold dilution is 7×10 4 TCID 50 / mL of virus has close to 100% neutralizing activity. After further humanization, it is expected to be developed into a neutralizing antibody drug for the treatment of diseases such as severe pneumonia caused by HADV-5 infection, filling the current lack of effective treatment options in this field and possessing significant scientific research value and clinical application prospects.

The invention discloses a novel coronavirus neutralizing antibody, a detection kit and application of the novel coronavirus neutralizing antibody. The amino acid sequence of a heavy chain variable region of the neutralizing antibody is shown as SEQ ID No.1, and the amino acid sequence of a light chain variable region of the neutralizing antibody is shown as SEQ ID No.2. The antibody with mature affinity is screened through bioinformatics analysis of a single B cell, the antibody screening process is optimized by combining single cell RNA sequencing, VDJ rearrangement analysis and somatic cell hypermutation research, blindness of a traditional method is avoided, and the accuracy and effectiveness of antibody screening are improved. According to the neutralizing antibody GR12 provided by the invention, a heavy chain variable region and a light chain variable region of the neutralizing antibody GR12 can be specifically combined with an RBD structural domain of the SARS-CoV-2 and an S-Trimer structural domain of an Omicro variant, so that a broad-spectrum neutralizing effect on the SARS-CoV-2 virus and the variant thereof is realized. The binding activity of the antibody GR12 to S-Trimer and RBD under 2-fold and 300-fold dilution conditions is obviously superior to that of other antibodies, which indicates that the antibody GR12 has high affinity and dilution stability and is suitable for clinical large-dose administration.

The invention discloses a novel coronavirus neutralizing antibody, a detection kit and application of the novel coronavirus neutralizing antibody. The amino acid sequence of a heavy chain variable region of the neutralizing antibody is shown as SEQ ID No.1, and the amino acid sequence of a light chain variable region of the neutralizing antibody is shown as SEQ ID No.2. The antibody with mature affinity is screened through bioinformatics analysis of a single B cell, the antibody screening process is optimized by combining single cell RNA sequencing, VDJ rearrangement analysis and somatic cell hypermutation research, blindness of a traditional method is avoided, and the accuracy and effectiveness of antibody screening are improved. According to the neutralizing antibody GR75 provided by the invention, a heavy chain variable region and a light chain variable region of the neutralizing antibody GR75 can be specifically combined with an RBD structural domain of the SARS-CoV-2 and an S-Trimer structural domain of an Omicro variant, so that a broad-spectrum neutralizing effect on the SARS-CoV-2 virus and the variant thereof is realized. The binding activity of the antibody GR75 to S-Trimer and RBD under 2-fold and 300-fold dilution conditions is obviously superior to that of other antibodies, which indicates that the antibody GR75 has high affinity and dilution stability and is suitable for clinical large-dose administration.

The invention discloses a novel coronavirus neutralizing antibody GR46, a detection kit and application of the novel coronavirus neutralizing antibody GR46. The amino acid sequence of a heavy chain variable region of the neutralizing antibody is shown as SEQ ID No.1, and the amino acid sequence of a light chain variable region of the neutralizing antibody is shown as SEQ ID No.2. The antibody with mature affinity is screened through bioinformatics analysis of a single B cell, the antibody screening process is optimized by combining single cell RNA sequencing, VDJ rearrangement analysis and somatic cell hypermutation research, blindness of a traditional method is avoided, and the accuracy and effectiveness of antibody screening are improved. According to the neutralizing antibody GR46 provided by the invention, a heavy chain variable region and a light chain variable region of the neutralizing antibody GR46 can be specifically combined with an RBD structural domain of the SARS-CoV-2 and an S-Trimer structural domain of an Omicro variant, so that a broad-spectrum neutralizing effect on the SARS-CoV-2 virus and the variant thereof is realized. The binding activity of the antibody GR46 to S-Trimer and RBD under 2-time and 300-time dilution conditions is obviously superior to that of other antibodies, which indicates that the antibody GR46 has high affinity and dilution stability and is suitable for clinical large-dose administration.

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 monoclonal antibody, a hybridoma cell secreting the monoclonal antibody and application of the hybridoma cell, and belongs to the field of peptides. The invention aims to provide a monoclonal antibody and a hybridoma cell secreting the monoclonal antibody. The hybridoma cell secreting the monoclonal antibody disclosed by the invention is a hybridoma cell 3G3, and the preservation number of the hybridoma cell 3G3 in the China General Microbiological Culture Collection Center (CGMCC) is CGMCC No.46345. The monoclonal antibody and the BVDV antibody detection kit based on the monoclonal antibody and the blocking ELISA method can be used for detecting a neutralizing antibody of the BVDV, and have the characteristics of strong specificity, good sensitivity, high sensitivity, high accuracy and the like.

The monoclonal antibody of the glycoprotein Gn of the severe fever with thrombocytopenia syndrome virus is screened on the basis of immune repertoire sequencing, and the biological characteristics of the monoclonal antibody are identified. The method comprises the following steps: taking purified SFTSV-Gn as bait, taking virus specific B cells from an immunized female BALB / c mouse B cell bank, selecting an antibody sequence with potential binding activity by utilizing a 10 * immune repertoire sequencing method in combination with bioinformatics analysis, constructing the antibody sequence to an eukaryotic expression vector pcDNA3.4, expressing and purifying a monoclonal antibody, and identifying the biological activity of the monoclonal antibody. 14 antibodies are selected from 3544 captured B cells, functional verification shows that 12 antibodies can be combined with SFTSV-Gn glycoprotein, and the Gn-5 antibody and the Gn-7 antibody have high combining capacity; western blot, indirect immunofluorescence test and flow cytometry prove that the antibody can specifically recognize the SFTSV, the effective concentration is as low as 2 mu g / mL, and a foundation is laid for development of a rapid diagnosis reagent for the SFTSV.

The invention discloses a cell membrane protein replacement type oncolytic virus vector and application thereof. The cell membrane protein replacement type oncolytic virus vector is rhabdoviridae virus, and a nucleotide sequence for coding G protein in a genome of the rhabdoviridae virus is replaced by a nucleotide sequence for coding an antibody and a nucleotide sequence for coding a spike protein truncation of coronavirus. The invention also discloses a construction method of the cell membrane protein replacement type oncolytic virus vector for expressing the antibody, the non-replicated virus vector is used for expressing the antibody sequence for the first time, and meanwhile, the novel coronavirus cell membrane protein is embedded into the virus surface, so that the cell membrane protein replacement type oncolytic virus vector can be rapidly produced in a suspension cell in a large scale; through removal of virus cell membrane protein genes, the virus cell membrane protein genes cannot be continuously replicated in vivo, so that the safety of the virus cell membrane protein genes is ensured, the tumor immunosuppression condition is improved, and an organism can be stimulated to generate a neutralizing antibody for resisting new coronavirus while tumor cells are killed.

The invention belongs to the technical field of biology, and particularly relates to a porcine epidemic diarrhea virus broad-spectrum neutralizing antibody and application thereof. A porcine epidemic diarrhea virus broad-spectrum neutralizing antibody 6G2 is screened, the virus neutralizing activity of the antibody is evaluated through a cell model, and it is found that the antibody can effectively block infection of G1a, G1b, G2a, G2b, G2c and S-INDEL (porcine epidemic diarrhea virus) gene subtype strains with high epidemic rate on cells. Further, animal infection experiments of pigs prove that the neutralizing antibody provided by the invention has good prevention and treatment effects on the porcine epidemic diarrhea virus, and has important significance on improving the capability of preventing and controlling porcine epidemic diarrhea in a pig farm.

The invention discloses an influenza A virus neutralizing antibody and application thereof, the neutralizing antibody comprises a heavy chain and a light chain, the heavy chain comprises a heavy chain CDR1 with an amino acid sequence as shown in SEQ ID NO: 5, a heavy chain CDR2 with an amino acid sequence as shown in SEQ ID NO: 6 and a heavy chain CDR3 with an amino acid sequence as shown in SEQ ID NO: 7; the light chain comprises a light chain CDR1 with an amino acid sequence as shown in SEQ ID NO: 8, a light chain CDR2 with an amino acid sequence as shown in SEQ ID NO: 9 and a light chain CDR3 with an amino acid sequence as shown in SEQ ID NO: 10. The antibody disclosed by the invention has high-efficiency broad-spectrum neutralizing activity in influenza A virus, and can be combined with various subtypes of hemagglutinin and inhibit splitting of hemagglutinin precursor protein and conformational change of the hemagglutinin in an acid environment. The invention has important significance and application prospect for preventing and treating diseases caused by influenza A virus.

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.

The invention provides a new bunyavirus neutralizing antibody screening method based on single cell BCR sequencing. The new bunyavirus neutralizing antibody screening method comprises the following steps: S1, collecting peripheral blood B cells of acute stage and recovery stage patients and healthy volunteers of new bunyavirus infected persons; s2, carrying out CD19 < + > CD38 < + > CD138 < + > magnetic bead separation to obtain a single-cell suspension; s3, constructing a single cell transcriptome and a BCR library, separating single cells by using a microfluidic system, and performing high-throughput sequencing; s4, analyzing a B cell subset, a BCR cloning amplification mode and a VJ gene pair enrichment characteristic through bioinformatics analysis; and S5, screening a specific BCR clone type related to the severity of the disease as a neutralizing antibody candidate target. The method provided by the invention provides powerful support for immune monitoring and vaccine design of new bunyavirus infection, and is expected to provide new strategies and methods for clinical treatment and prevention.

The invention relates to a method for stimulating an immune response by intramuscular injection of an artificial nucleic acid molecule comprising an open reading frame encoding an antigen and a 3′-UTR comprising at least two poly(A) sequences. The method may yield an increased immune response to the antigen or an increased neutralizing antibody response to the antigen.

Highly potent respiratory syncytial virus (RSV) neutralizing monoclonal antibodies that specifically bind RSV fusion protein (F) or RSV attachment glycoprotein (G) are described. The monoclonal antibodies were synthesized by sequencing RSV G- and RSV F-specific B cells from a human participant of an RSV challenge study. The RSV-specific monoclonal antibodies described herein exhibit potent neutralization of RSV (IC50 as low as 0.4 ng / ml) that is superior to previously disclosed RSV neutralizing antibodies. The RSV-specific monoclonal antibodies can be used, for example, to treat, inhibit and detect RSV infection.

In various embodiments immunogenic nanoparticles are provided that are capable of raising an immune response directed against SARS-CoV-2. In certain embodiments the immunogenic nanoparticles comprise mRNA multi-epitope vaccines that can be used in combination with or independent of other covid-19 vaccines (e.g., the spike protein mRNA vaccine(s)) to invoke a strong CD8+ or CD4+ T-cell as well as neutralizing antibody producing B-cell responses. In certain embodiments this vaccine is based on the rational combination of well-conserved T- and B-cell epitopes identified COVID-19 and viral variants.

The invention relates to a novel three-antigen HSV-2 subunit vaccine as well as a preparation method and application thereof, and belongs to the technical field of biology. The novel three-antigen HSV-2 subunit vaccine comprises antigens and a composite adjuvant, the antigens comprise HSV-2 gB2 envelope glycoprotein, HSV-2 gC2 envelope glycoprotein and HSV-2 gD2 envelope glycoprotein, and the composite adjuvant is CpG oligonucleotide and an aluminum adjuvant; the vaccine provided by the invention can induce a high-level neutralizing antibody, and widens targets for developing a novel multi-target antigen HSV-2 virus vaccine.

The invention discloses application of a neutralizing antibody of oncostatin M in preparation of a medicine for treating bronchial pulmonary dysplasia. The application proves that the intrauterine inflammation can cause BPD-like lung lesion to the filial generation mouse, and the OSM expression level in the mouse lung tissue one day after the intrauterine inflammation filial generation is grown is obviously higher than that of a control group; the OSM level of the BPD group is higher than that of the non-BPD group in the umbilical blood sample of the premature infant; in-vitro experiments show that the OSM treatment can cause lung epithelial cell injury; by dripping a recombined OSM neutralizing antibody into the nose of a newborn mouse, down-regulation of the OSM level in the lung of the mouse is realized, filial generation lung injury induced by the intrauterine inflammation can be repaired, and the OSM can be used as a treatment target of the BPD caused by the intrauterine inflammation.

The invention discloses a vaccine composition and a preparation method thereof, the vaccine composition comprises a first component and a second component, the first component is a recombinant protein antigen, the second component is an oil-in-water composite adjuvant, and the oil-in-water composite adjuvant comprises a water phase, an oil phase, an emulsifier, a Toll-like receptor stimulant and / or an STING stimulant. The vaccine composition provided by the invention can stimulate a body to generate a high-level specific IgG antibody and a neutralizing antibody, and has a good cellular immune effect at the same time. The second component provided by the invention is simple in preparation process, low in cost, stable in dosage form, easy to store and good in immune effect, and has a wide application prospect in vaccine or drug development.

The invention provides a PDCoV virus mRNA (messenger Ribonucleic Acid) vaccine capable of self-cleaving and expressing a plurality of virus structural proteins and a preparation method of the PDCoV virus mRNA vaccine, and the vaccine comprises mRNA for expressing S, M and N proteins of a PDCoV virus and LNP for encapsulating the mRNA, and the LNP is marked as SMN-mRNA-LNP. The invention provides a PDCoV mRNA vaccine strategy based on combination of S, M and N for the first time, the S, M and N structural proteins of the PDCoV are connected by using a self-cleavage peptide P2A, the S protein is subjected to double proline mutation, so that a single mRNA can express multiple PDCoV antigens, and a multi-level defense system is constructed by using the neutralizing antibody induction capability of the S protein, the immune regulation function of the M protein and the cellular immune activation characteristic of the N protein. Through evaluation of immunogenicity, antibody level and challenge protection effect of the vaccine in mice, suckling piglets and pregnant sows, a new idea is provided for development of broad-spectrum and efficient PDCoV vaccines, and a practical basis is provided for research and development of coronavirus multi-antigen mRNA vaccines.

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 discloses a porcine reproductive and respiratory syndrome virus RNA vaccine and application thereof, and relates to the technical field of veterinary biological products. According to the present invention, the coding sequence of the porcine reproductive and respiratory syndrome virus GP5 protein is optimized, and the GP5 protein is connected with the M protein through the connecting peptide, such that the saRNA prepared by using the nucleic acid molecule can produce the efficient expression on the GP5 protein and the M protein after the cell transfection. After the optimized porcine reproductive and respiratory syndrome GP5 protein and M protein fusion protein is immunized, the level of neutralizing antibodies generated by animals can be improved, and the effect of preventing the porcine reproductive and respiratory syndrome virus is achieved. Therefore, the nucleic acid molecule provided by the invention can be used for developing or preparing nucleic acid vaccines. Compared with a traditional vaccine, the vaccine provided by the invention has good immunogenicity and safety, and has a good application prospect.

The invention discloses a culture medium for A type clostridium perfringens and C type clostridium perfringens, a fermentation culture method and application of fermentation toxin production. Wherein the culture medium comprises the following components: tryptone, # imgabs0 # peptone, casein peptone, a yeast extract, lactoalbumin hydrolysate, trihydroxymethyl aminomethane, sodium acetate and glucose. According to the present invention, through the specific formula composition, the A-type clostridium perfringens and the C-type clostridium perfringens can adapt to fermentation in the fermentation tank, and in the fermentation culture process, the toxin production ability of the A-type clostridium perfringens and the C-type clostridium perfringens can be significantly improved by controlling the stirring speed, the pH value of the fermentation liquid and the precise material supplementing manner. And the supernate obtained by fermentation is used as a raw material for preparing the vaccine, so that animals can generate high-level neutralizing antibodies aiming at the clostridium perfringens A and C type toxins after being immunized with the vaccine, the death of the animals caused by infection of the clostridium perfringens A and C type toxins is avoided, and the method has practical popularization and application values.

The invention provides a method for detecting an anti-C1 esterase inhibitor neutralizing antibody in a biological sample, and the method mainly utilizes the characteristic that protein A resin is combined with an Fc segment of the antibody to capture the antibody (containing the anti-C1 esterase inhibitor neutralizing antibody), so that the C1 esterase inhibitor and complement component 1s (C1s) in the biological sample after thermal inactivation are removed; furthermore, an antibody (containing a neutralizing antibody) or an anti-C1 esterase inhibitor compound on the protein A resin is subjected to acid dissociation through an acidolysis solution, so that not only can the anti-C1 esterase inhibitor neutralizing antibody existing in a free form in a biological sample be detected, but also the anti-C1 esterase inhibitor neutralizing antibody existing in a combined form in the biological sample can be detected.

An antibody against respiratory syncytial virus (RSV) and / or human metapneumovirus (HMPV) is described herein. The antibody includes binding domains from anti-idiotypic monoclonal antibodies (ai-mAbs) that bind an RSV neutralizing antibody VH3-21 / VL1-40 presented as B cell receptors (BCRs). The antibodies can be used as a vaccine to selectively elicit antibodies capable of neutralizing RSV and / or HMPV without a need for somatic mutation. The vaccines are particularly useful to treat and / or reduce the risk of RSV and / or HMPV infection in infants.

The invention provides a porcine epidemic diarrhea virus G2c subtype strain and application thereof. According to the invention, the porcine epidemic diarrhea virus G2c subtype HN2501 strain is separated from porcine intestinal tissues and is subjected to passage purification, and the microbial preservation number of the porcine epidemic diarrhea virus G2c subtype HN2501 strain is CGMCC (China General Microbiological Culture Collection Center) No.46417. The isolated strain can stably proliferate on passage cells to generate typical cytopathy. The G2c subtype isolated strain of the porcine epidemic diarrhea virus has excellent immunogenicity; the vaccine prepared from the isolated strain can induce piglets to generate high-level neutralizing antibodies; meanwhile, the ELISA kit prepared from the isolated strain can be used for well detecting the porcine epidemic diarrhea virus antibody in serum.

The invention provides a subunit vaccine composition for porcine epidemic diarrhea, porcine delta coronal and porcine rotavirus as well as a preparation method and application of the subunit vaccine composition, and is characterized in that the subunit vaccine composition comprises prokaryotically expressed swine erysipelas filamentous bacillus SpaA protein, viral subunit protein and pharmaceutically acceptable adjuvants; wherein the amino acid sequence of the prokaryotically expressed swine erysipelas filamentous bacillus SpaA protein is as shown in SEQ No.1, and the viral subunit protein is selected from one or more of porcine epidemic diarrhea virus S protein, porcine delta coronavirus S protein and porcine rotavirus VP8 protein. The vaccine composition disclosed by the invention has the advantages of strong immunogenicity, good safety, no immune interference, high neutralizing antibody titer and long antibody duration.