41 results about "Virus Neutralization" patented technology

The invention discloses a chikungunya virus envelope E2 protein monoclonal antibody and application thereof, and belongs to the technical field of medicines. The chikungunya virus envelope protein E2 expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the chikungunya virus envelope protein E2 are screened from rabbit spleen cells through flow sorting, and signal peptide and variable region gene fragments of an antibody are cloned through reverse transcription-polymerase chain reaction. According to the present invention, the chikungunya virus-resistant monoclonal antibody with high neutralizing activity and capable of 100% protection of mice against chikungunya virus lethal attack is obtained by carrying out enzyme-linked immunosorbent assay, in-vitro virus neutralization and mouse toxicity attack experiment after mammalian cell expression and purification, and the recombinant chikungunya virus-resistant monoclonal antibody has characteristics of high neutralizing activity and high immunogenicity, and can be used for preparing the chikungunya virus-resistant monoclonal antibody, and the recombinant chikungunya virus-resistant monoclonal antibody. The monoclonal antibody has application value in prevention and treatment of yellow fever.

Isolated monoclonal antibodies and antigen binding fragments thereof are disclosed that specifically binds to West Nile Virus. These monoclonal antibodies or antigen binding fragments can inhibit a West Nile Virus infection in a subject, and / or can be used to diagnose a West Nile Virus in a subject. Nucleic acid molecules encoding these antibodies and antigen binding fragments are also disclosed, as are vectors including these nucleic acid molecules, and host cells expressing these vectors.

This invention belongs to the field of biotechnology, specifically relating to a broad-spectrum neutralizing antibody against porcine epidemic diarrhea virus (PEDV) and its applications. This invention screened a broad-spectrum neutralizing antibody against PEDV, 6G2. Cell model evaluation revealed that the antibody effectively blocked infection of cells by several highly prevalent PEDV genotypes: G1a, G1b, G2a, G2b, G2c, and S-INDEL. Further animal infection experiments in pigs demonstrated that the neutralizing antibody provided by this invention has good preventive and therapeutic effects against PEDV, which is of great significance for improving the ability of pig farms to control PEDV.

The invention discloses a yellow fever virus envelope protein monoclonal antibody and application thereof, and belongs to the technical field of medicines. Yellow fever virus envelope protein E expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the yellow fever virus envelope protein E are screened from rabbit spleen cells through flow sorting, and signal peptide and variable region gene fragments of an antibody are cloned through reverse transcription-polymerase chain reaction. According to the present invention, the monoclonal antibody with high neutralizing activity and capable of completely protecting mice from yellow fever virus lethal attack is obtained by using the monoclonal antibody as a template, connecting the monoclonal antibody and the constant region gene to an expression vector, carrying out mammalian cell expression and purification, and carrying out enzyme-linked immunosorbent assay, in-vitro virus neutralization and mouse virus attack experiment, such that the monoclonal antibody has high neutralizing activity and can completely protect mice from yellow fever virus lethal attack; the monoclonal antibody has application value in prevention and treatment of yellow fever.

This invention relates to the field of biotechnology, and in particular to an antigenic epitope peptide, product, and application for detecting antibodies produced by porcine deltacoronavirus infection. This invention identified short peptide epitopes in PDCoV longitudinal challenge serum samples, obtaining 12 short peptides, which were then prepared into a porcine deltacoronavirus antibody detection kit. Finally, six field serum samples were selected for antibody detection, and the effectiveness was verified using a virus neutralization experiment. The results showed that the porcine deltacoronavirus antibody detection kit provided by this invention is effective. Furthermore, the porcine deltacoronavirus antibody detection kit provided by this invention, as a novel detection method, improves sensitivity and specificity while simultaneously providing multiple detection results, namely, determining whether viral infection has occurred, identifying the stage of infection, and achieving DIVA (Differentiation, Infection, and Variability) analysis.

The application provides a preparation method of a shrimp iridovirus DIV1 neutralizing antibody and application thereof. The gene sequence shown as SEQ ID NO. 1 and the amino acid sequence shown as SEQ ID NO. 2 are used to solve the problem that there is no neutralizing antibody for the shrimp iridovirus DIV1 at present. The antibody for the shrimp iridovirus DIV1 prepared by the application can specifically recognize the DIVI virus and has good virus neutralizing activity, can effectively reduce the death of shrimps caused by DIV1 virus infection, significantly improve the ability of shrimps to resist DIV1 virus infection, and effectively prevent the infection and transmission of the virus.

The invention discloses a cat parvovirus VP2 protein nano antibody as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The amino acid sequence of the cat parvovirus VP2 protein nano antibody is as shown in SEQ ID NO. 1 or SEQ ID NO. 2. According to the invention, an alpaca is immunized by using FPV VP2 protein, a phage display nano antibody library is constructed, and two nano antibodies 3D6 and 1B9 with strong specificity and high affinity are obtained through multiple rounds of panning and identification. Through identification, the nano antibodies 3D6 and 1B9 can be specifically combined with FPV and VP2 proteins, and have high affinity and remarkable virus neutralizing activity. The nano antibody can be used for preparing a detection reagent or kit for feline parvovirus infection, and rapid and accurate diagnosis is realized; the compound can also be used as an active component to prepare a therapeutic drug, and provides a special therapeutic scheme for FPV infection. The nano antibody provides an important material basis for developing a novel diagnostic tool and a specific therapeutic drug aiming at the feline parvovirus.

The invention discloses an anti-Crimean-Congo hemorrhagic fever virus (CCHFV) neutralizing antibody N013 and application thereof in diagnosis, treatment and / or prevention of Crimean-Congo hemorrhagic fever, and the antibody has a unique CDR region and can be combined with the Gc protein of the CCHFV with high affinity. In a virus neutralization experiment, the antibody N013 disclosed by the invention has strong neutralization activity on CCHFV pseudovirus particles, the IC50 value is 26.63 ng / mL, and the antibody N013 has a wide application prospect in treatment of Crimean-Congo hemorrhagic fever.

The invention discloses a chikungunya virus monoclonal antibody and application thereof in preparation of preventive and therapeutic antibody drugs, and belongs to the technical field of medicines. The chikungunya virus envelope protein E2 expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the chikungunya virus envelope protein E2 are screened from rabbit spleen cells through flow sorting, and signal peptide and variable region gene fragments of an antibody are cloned through reverse transcription-polymerase chain reaction. According to the present invention, the chikungunya virus-resistant monoclonal antibody with high neutralizing activity and capable of 100% protection of mice against chikungunya virus lethal attack is obtained by carrying out enzyme-linked immunosorbent assay, in-vitro virus neutralization and mouse toxicity attack experiment after mammalian cell expression and purification, and the recombinant chikungunya virus-resistant monoclonal antibody has characteristics of high neutralizing activity and high immunogenicity, and can be used for preparing the chikungunya virus-resistant monoclonal antibody, and the recombinant chikungunya virus-resistant monoclonal antibody. The monoclonal antibody has application value in prevention and treatment of yellow fever.

The application belongs to the field of molecular biology and particularly relates to an anti-CD4 nanobody with a longer plasma half-life, which comprises a CD4 targeting domain and an HSA targeting domain; the CD4 targeting domain comprises three complementarity determining regions CDR1-3, and the sequences are shown in SEQ ID NO:1-3 in sequence; the HSA targeting domain comprises three complementarity determining regions CDR1-3, and the sequences are shown in SEQ ID NO:4-6 in sequence. The application develops a nanobody drug for attacking the target point CD4 of HIV, and constructs a "Ushanka" type nanobody NbCD4-NbHSA-NbCD4 by introducing an anti-HSA VHH, so that the half-life of the nanobody can be effectively improved; meanwhile, the efficacy of the single nanobody and the "Ushanka" type nanobody in treating HIV infection is evaluated by using a pseudovirus neutralization experiment. The application provides a potential effective nanobody new drug for the clinical treatment of HIV.

The present application relates to a kind of anti-procambarus clarkii white spot syndrome virus monoclonal antibody and its application, the complementary determining region CDR1-3 sequence of the heavy chain variable region of the anti-procambarus clarkii white spot syndrome virus monoclonal antibody is sequentially shown as SEQ ID NO:3-5, the complementary determining region CDR1-3 sequence of the light chain variable region is sequentially shown as SEQ ID NO:6-8.The present application successfully obtains the high affinity of anti-VP28 monoclonal antibody, and it has excellent affinity to procambarus clarkii white spot syndrome virus.The monoclonal antibody can be used for specific detection to procambarus clarkii white spot syndrome virus, and has good virus neutralization activity.By adding the monoclonal antibody in water body, the resistance of procambarus clarkii to white spot syndrome virus can be greatly improved, easy to operate, without injection, suitable for large-scale breeding scene, provide a new technical means for the prevention and control of white spot syndrome virus disease in aquaculture.

The application provides two kinds of anti-rabies virus full human monoclonal antibodies or a combination thereof, a heavy chain variable region of the antibodies comprises three CDR regions, sequences of the three CDR regions are respectively shown as SEQ ID NO:1-3 and SEQ ID NO:9-11, and a light chain variable region of the antibodies comprises three CDR regions, sequences of the three CDR regions are respectively shown as SEQ ID NO:4-6 and SEQ ID NO:12-14. The application also provides application of the antibodies to anti-rabies virus. The antibodies are high in affinity to different antigen epitopes of a rabies virus G protein, and are high in potency in neutralizing rabies virus strains derived from different genetic subtypes, and the combination of the antibodies does not affect antigen binding and virus neutralization activities of the antibodies.

The present application relates to a kind of anti-procambarus clarkii white spot syndrome virus monoclonal antibody and its application, the complementary determining region CDR1-3 sequence of the heavy chain variable region of the anti-procambarus clarkii white spot syndrome virus monoclonal antibody is sequentially shown as SEQ ID NO:3-5, the complementary determining region CDR1-3 sequence of the light chain variable region is sequentially shown as SEQ ID NO:6-8.The present application successfully obtains the high affinity of anti-VP28 monoclonal antibody, and it has excellent affinity to procambarus clarkii white spot syndrome virus.The monoclonal antibody can be used for specific detection to procambarus clarkii white spot syndrome virus, and has good virus neutralization activity.By adding the monoclonal antibody in water body, the resistance of procambarus clarkii to white spot syndrome virus can be greatly improved, easy to operate, without injection, suitable for large-scale breeding scene, provide a new technical means for the prevention and control of white spot syndrome virus disease in aquaculture.

The invention relates to a multivalent vaccine, in particular to a chimera multivalent vaccine based on Ercovirus type 11, 18 and 30 key epitopes as well as a preparation method and application of the chimera multivalent vaccine. According to the invention, a chimera multivalent vaccine VP1-III is also designed based on key epitopes, an E11-VP1 sequence is taken as a basis, one E18-VP1 key epitope and two E30-VP1 key epitopes are respectively inserted into the E11-VP1 sequence to obtain a chimera multivalent protein vaccine VP1-III, and the chimera multivalent protein vaccine VP1-III is taken as an immunogen to immunize a mouse to detect antibody titer and virus neutralizing capacity of antiserum. A new thought is provided for designing and preparing multivalent vaccines and detecting immunogenicity of viral vaccines.

The application discloses an anti-Western Nile virus neutralizing antibody WN-XH3 and application thereof. The antibody comprises a heavy chain variable region and a light chain variable region, has good binding activity with Western Nile virus E protein, and has strong virus neutralizing activity, thereby laying a foundation for diagnosis and treatment of Western Nile virus infection related diseases, development of a Western Nile virus infection related disease treatment drug and the like, and can be used as a candidate of a clinical anti-Western Nile virus drug, and has a good application prospect.

The application discloses a dual-epitope llama-derived nanobody and application thereof, and belongs to the field of biological medicine. The application is aimed at different epitopes of MERS-CoV, and two strains of antibodies 2-77 and 4-58 aiming at different binding sites of MERS-CoV RBD are obtained by immunizing a llama, performing specific screening on a ribosome display nanobody library, performing a pseudovirus neutralization experiment, performing a competitive BLI experiment, identifying VHH epitopes and molecular docking by using a phage random peptide library, and constructing a dual-epitope llama-derived nanobody 2F4 with good neutralization activity, high expression and stability by using the two strains of nanobodies. It is proved by a lesion formation test and a mouse in-vivo experiment that the dual-epitope llama-derived nanobody 2F4 constructed by the application can effectively neutralize MERS-CoV and effectively protect animals, and the application provides a train of thought and technical support for exploring new antigen epitopes of MERS-CoV and designing a therapeutic antibody drug.

The invention discloses a chikungunya virus envelope E2 protein monoclonal antibody and application of the chikungunya virus envelope E2 protein monoclonal antibody in preparation of preventive and therapeutic antibody drugs, and belongs to the technical field of medicines. The chikungunya virus envelope protein E2 expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the chikungunya virus envelope protein E2 are screened from rabbit spleen cells through flow sorting, and signal peptide and variable region gene fragments of an antibody are cloned through reverse transcription-polymerase chain reaction. According to the present invention, the chikungunya virus-resistant monoclonal antibody with high neutralizing activity and capable of 100% protection of mice against chikungunya virus lethal attack is obtained by carrying out enzyme-linked immunosorbent assay, in-vitro virus neutralization and mouse toxicity attack experiment after mammalian cell expression and purification, and the recombinant chikungunya virus-resistant monoclonal antibody has characteristics of high neutralizing activity and high immunogenicity, and can be used for preparing the chikungunya virus-resistant monoclonal antibody, and the recombinant chikungunya virus-resistant monoclonal antibody. The monoclonal antibody has application value in prevention and treatment of yellow fever.

The application belongs to the technical field of biological medicine, and provides a DNA programmed virus neutralizing multivalent aptamer medicament and a preparation method thereof.The medicament comprises a circular DNA as a template, and a DNA multivalent neutralizing aptamer particle is synthesized through a rolling circle amplification reaction, wherein the DNA template can be designed to be directed against a plurality of virus antigen epitope neutralizing aptamers.The DNA programmed virus neutralizing multivalent aptamer medicament provided by the application is simple to operate, raw materials are easy to obtain, has good biocompatibility, and has a wide application range, can effectively inhibit the invasion of virus particles into the cell interior, can capture and close the virus particles, thereby inhibiting the infection of the virus particles on the host cells, and can play a safe and effective virus neutralizing role and block the transmission and infection of the virus.

The invention discloses an akabane disease virus monoclonal antibody with virus neutralizing activity and a colloidal gold immunochromatography test strip prepared from the akabane disease virus monoclonal antibody, and belongs to the technical field of biology. The monoclonal antibody is secreted by a hybridoma cell strain with the preservation number of CGMCC (China General Microbiological Culture Collection Center) NO.45859, and is named as 1H6. The monoclonal antibody provided by the invention can specifically recognize AKAV N protein and can neutralize akabane disease virus. A purified monoclonal antibody 1H6 is coupled with gold particles, a rabbit anti-AKAV polyclonal antibody is coated on an NC membrane to serve as a detection T line, a goat anti-mouse IgG antibody is coated on an NC membrane to serve as a quality control C line, the colloidal gold immunochromatography test strip for detecting akabane disease viruses is constructed, and the colloidal gold immunochromatography test strip can be used for detecting akabane disease virus antigens. The method has the advantages of no need of professionals, simplicity in operation, high detection sensitivity and simple and convenient operation requirements, can meet rapid and mass screening detection of grassroots enterprises and public institutions, and can also be used for detection analysis such as scientific research and the like.

The invention discloses a nano antibody targeting rabies virus G protein and application of the nano antibody, and belongs to the technical field of biological medicine. A heavy chain variable region of the nano antibody comprises three complementary determining regions CDR1, CDR2 and CDR3; wherein the amino acid sequence of the CDR1 is as shown in SEQ ID NO. 1, the amino acid sequence of the CDR2 is as shown in SEQ ID NO. 2, and the amino acid sequence of the CDR3 is as shown in SEQ ID NO. 3. The antibody obtained by the invention has high specific recognition and binding capacity to G protein, has nanomole affinity to target antigen, has good application prospect in preparation of drugs for preventing and / or treating rabies, has the characteristics of high purity, low endotoxin and amplified production, and can be used for preparing drugs for preventing and / or treating rabies. By coupling with RVG29 and NIR-Pep, the problems that the antibody cannot penetrate through a blood brain barrier plate and the endosome escape rate is low can be solved, and the intracellular virus neutralization efficiency is improved.

This invention discloses an anti-Dabie Banda virus Gn protein nanobody, a bispecific antibody, and their applications, belonging to the field of biodetection technology. This invention constructs an alpaca natural nanobody library, using recombinantly expressed Gn protein as the antigen. Through multiple rounds of gradient screening using phage display technology, three nanobodies—G15, G20, and G40—that specifically bind to the Dabie Banda virus Gn protein are obtained. All three antibodies exhibit excellent antigen-binding specificity and virus-neutralizing activity; among them, G40 shows the most prominent neutralizing effect. The bispecific antibody G40-C1q, constructed based on G40, combines Gn protein and human C1q protein, achieving a synergistic effect of antiviral activity and inhibition of inflammatory damage. It can be used for the development of drugs for the prevention and treatment of Dabie Banda virus infection and for high-specificity detection reagents, providing candidate antibody drugs for the treatment of fever with thrombocytopenia syndrome, and providing a reliable tool for virus-related research, disease diagnosis, and clinical translation.

The application provides a polypeptide molecule for specifically recognizing HIV gp120 protein and an application thereof, and belongs to the technical field of biological medicine. The application provides a lead polypeptide derivative for specifically targeting and combining HIV gp120, wherein the lead polypeptide comprises an amino acid sequence shown in SEQ ID No. 1. The application also provides a lead polypeptide derivative prepared based on the lead polypeptide, and a polypeptide nanoparticle constructed by the lead polypeptide derivative, and both of them have gp120 protein combining capacity. The derivative and the nanoparticle have good virus neutralization inhibition effect. The series of polypeptide molecules can specifically combine with free HIV virus, HIV infected cells and free gp120, so as to block HIV infected cells, delay the AIDS disease process, realize AIDS treatment drug targeted delivery, detection and diagnosis.

The application discloses an enhanced CD46 decoy protein and application thereof in preparation of a medicine against bovine viral diarrhea virus, and belongs to the technical field of biological medicine. The application first analyzes the crystal structure of an E2-CD46 complex, determines key amino acid sites of a binding interface between E2 and CD46, and constructs a mutant with significantly improved affinity compared with a wild type. The application further constructs a dimer and a trimer with a CD46 truncated body or mutant as a monomer. The obtained high-affinity decoy proteins mCD46-12 and mCD46-13 can achieve nearly complete virus neutralization and blocking effect at micromolar concentration, can effectively competitively inhibit invasion of BVDV to host cells, provide a brand-new and efficient candidate molecule for preparation of a medicine for treating bovine viral diarrhea infection, and have great clinical application prospect and economic and social value.

This invention discloses a method for screening trace amounts of neutralizing antibodies against canine distemper virus (CDV). The method involves neutralizing the sample with live canine distemper virus, then inoculating it into Vero-Slam cells, using fluorescent lesions as the criterion. Detecting neutralizing antibodies using the true conformation of the virus ensures the highest degree of accuracy in detecting neutralizing antibodies. CDV-positive neutralizing serum (neutralizing titer 1:708) was serially diluted twofold to multiple concentration groups as the sample to be tested. The detection rate and limit of detection of CDV neutralizing activity of the method for each dilution of serum were determined. Results showed that serum with a CDV neutralizing titer of 1:708 diluted twofold... 12 The neutralizing activity after doubling is the lowest detection limit of the method of this invention, with a detection rate of 63.54 ± 4.774%. The method of this invention uses few reagents, none of which are expensive, resulting in low cost. This invention provides a new technical means for screening trace neutralizing antibodies against canine distemper virus.

The present invention discloses an anti-Crimean-Congo hemorrhagic fever virus (CCHFV) neutralizing antibody N015 and its use in the diagnosis, treatment and / or prevention of Crimean-Congo hemorrhagic fever, the antibody having a unique CDR region capable of binding to the Gc protein of CCHFV with high affinity. In a virus neutralization experiment, the antibody N015 disclosed by the invention has strong neutralization activity on CCHFV pseudovirus particles, the IC50 value is 53.39 ng / mL, and the antibody N015 has a wide application prospect in treatment of Crimean-Congo hemorrhagic fever.

The name of the invention: "Neutralization of viruses through oxidation-reduction reaction on the virus envelope" suggests its use in medicine, both in the human and animal fields. This is not only a specific use, but also from the point of view of the dosage regimen, the method of dosage, a new group of patients, of component No. 1 - Cu2+ obtained in a solution of copper sulfate pentahydrate in distilled water, where copper must have at least two free ions and component No. 2 - L-ascorbic acid for neutralization of viruses through oxidation-reduction reaction on the virus envelope, which is characterized by the fact that it ensures the destruction of the virus envelope and thus the inhibition of viruses in humans and other warm-blooded animals. Both substances are administered in a non-toxic concentration appropriate for the human and animal organism and with an appropriate time interval, so that the most suitable conditions for the oxidation-reduction reaction on the virus envelope are created and maintained. The use of these two components establishes a scientifically proven premise for the successful prevention and treatment of infection with various types of enveloped viruses and the diseases they cause.

The invention is applicable to the technical field of veterinary biomedicine, and provides a nano-antibody specifically bound with porcine circovirus type 2 and application thereof, nucleotide sequences of the nano-antibody are respectively shown as SEQ ID NO: 1 and SEQ ID NO: 2, and amino acid sequences of the nano-antibody are respectively shown as SEQ ID NO: 3 and SEQ ID NO: 4. The nano antibody provided by the invention can be specifically combined with porcine circovirus particles, virus neutralizing activity is shown in PK15 cells, and the multivalent nano antibody obtained by using a ferritin coupling system can significantly reduce PCV2 virus particle infection and replication and proliferation capacities.

This invention discloses an anti-Dabie Banda virus Gn protein nanobody, a bispecific antibody, and their applications, belonging to the field of biodetection technology. This invention constructs an alpaca natural nanobody library, using recombinantly expressed Gn protein as the antigen. Through multiple rounds of gradient screening using phage display technology, three nanobodies—G15, G20, and G40—that specifically bind to the Dabie Banda virus Gn protein are obtained. All three antibodies exhibit excellent antigen-binding specificity and virus-neutralizing activity; among them, G40 shows the most prominent neutralizing effect. The bispecific antibody G40-C1q, constructed based on G40, combines Gn protein and human C1q protein, achieving a synergistic effect of antiviral activity and inhibition of inflammatory damage. It can be used for the development of drugs for the prevention and treatment of Dabie Banda virus infection and for high-specificity detection reagents, providing candidate antibody drugs for the treatment of fever with thrombocytopenia syndrome, and providing a reliable tool for virus-related research, disease diagnosis, and clinical translation.

An SARS-COV-2 recombinant Spike protein is provided. The research process of the protein is as follows: the dominant strain in circulation was identified by screening clinical samples of SARS-COV-2 patients and its mutations in Turkey were evaluated by sequencing the Spike gene. Sequencing data and in silico methods were used to design the Spike antigen and then the novel Spike antigen was docked with the human ACE2 (Angiotensin Converting Enzyme-2) receptor to determine the binding energy. After DNA vaccine construction, HEK293T cells were transfected and analyzed for protein expression capacity by IFA, Western blot and RT-qPCR, then BALB / c mice and K18-hACE2 transgenic mice were immunized with DNA vaccine administered intramuscularly (IM) and intradermally (ID) using an electroporator device three times on days 0, 14 and 56. Humoral and cellular immune responses were then analyzed using recombinant ELISA, Western blot, surrogate virus neutralization assay, microneutralization assay, Cytokine ELISA and flow cytometry.