223 results about "Neutralization" patented technology

The present application relates generally to the field of biochemical detection. The invention provides a method and a kit for detecting an anti-IgE antibody in a biological sample. The method for detecting the anti-IgE antibody in the biological sample comprises the following steps: (a) contacting the biological sample with an acidolysis solution to dissociate the anti-IgE antibody from IgE so as to obtain an acidolysis sample; (b) contacting the acidolysis sample with a neutralization buffer solution to terminate acidolysis so as to obtain a to-be-detected sample; and (c) determining the anti-IgE antibody in the sample to be detected. The kit for detecting the anti-IgE antibody in the biological sample comprises an acidolysis solution and a neutralization buffer solution. The problem of background IgE interference in anti-IgE antibody detection is solved to a certain extent.

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 discloses a broad-spectrum monoclonal antibody capable of neutralizing bovine viral diarrhea virus and application of the broad-spectrum monoclonal antibody. The invention relates to the field of biological medicine, and provides an antibody which comprises a heavy chain variable region and a light chain variable region, the amino acid sequences of HCDR1, HCDR2 and HCDR3 in the heavy chain variable region are sequentially shown as the 26th site to the 33rd site, the 51st site to the 58th site and the 97th site to the 109th site of SEQ ID NO: 1; the amino acid sequences of the LCDR1, the LCDR2 and the LCDR3 in the light chain variable region are sequentially shown as the 27 to 32 sites, the 50 to 52 sites and the 89 to 99 sites of SEQ ID NO: 3. The antibody provided by the invention is a BVDV broad-spectrum neutralizing monoclonal antibody, can be used for BVDV antigen diagnosis and vaccine immune efficacy evaluation, and has important significance in prevention and control of BVD epidemic.

The invention discloses an anti-parvovirus canine-mouse chimeric antibody, and belongs to the field of gene engineering, the chimeric antibody comprises a heavy chain variable region, a light chain variable region, a heavy chain constant region and a light chain constant region, the heavy chain variable region and the light chain variable region are mouse-derived fragments, and the heavy chain constant region and the light chain constant region are canine-derived fragments; the amino acid sequence of the heavy chain variable region is as shown in SEQ ID No.2, and the amino acid sequence of the light chain variable region is as shown in SEQ ID No.4. The canine-mouse chimeric antibody has broad-spectrum high neutralization activity on various subtypes of CPV in vitro, the immunological rejection of a mouse antibody in a canine body is reduced, and the canine infected by CPV-2c can be effectively treated by using the canine-mouse chimeric antibody.

The present application provides a detection method and use for neutralizing antibodies for therapeutic monoclonal antibodies, which is a CLB NAb assay developed through target-based drug depletion and drug-based NAb extraction for CMAB009 with high drug and target tolerance. Acid is typically used in the pretreatment of a sample to dissociate the drug / NAb complex. In order to improve the treatment effect, a two-step method involving an amphiphilic method and a bead method is adopted. Initially, EGFR-coated magnetic beads (target beads) are used to eliminate the majority of free drugs in a sample. Subsequently, after acid dissociation of the sample, NAb is captured with beads. NAb is neutralized and eluted by acid, and a biotinylated drug is added into an EGFR-coated flat plate for detecting NAb. A comprehensive experiment strategy is implemented, meanwhile, drug interference is relieved, and target tolerance is enhanced. Through troubleshooting and optimization, NAb determination is verified to be used for clinical sample analysis, so that the NAb is successfully applied to III-stage clinical immunogenicity NAb detection of CMAB009. The result shows that the integrated detection optimization strategy can greatly improve the key performance characteristics of CLB NAb detection, so that the potential immunogenicity and the influence of the potential immunogenicity on better biological development can be evaluated more reliably.

The invention discloses an anti-respiratory syncytial virus antibody and application thereof. The anti-respiratory syncytial virus antibody or the antigen binding fragment thereof can specifically recognize the pre-F protein of the respiratory syncytial virus and neutralize the respiratory syncytial virus, especially has efficient neutralizing activity on an A2 strain of the respiratory syncytial virus, can effectively prevent and control infection of the respiratory syncytial virus, and can be used for preparing the anti-respiratory syncytial virus antibody or the antigen binding fragment thereof. A new thought is provided for detection of the respiratory syncytial virus and prevention and treatment of diseases related to respiratory syncytial virus infection, and wide application prospects are achieved.

The present invention discloses a combined monoclonal antibody preparation against rabies virus, which relates to the technical field of biomedicine. The combined monoclonal antibody preparation against rabies virus contains R92 antibody and R71 antibody; the light chain sequence of the R92 antibody is as shown in SEQ ID NO:1, the heavy chain sequence of the R92 antibody is as shown in SEQ ID NO:2, the light chain sequence of the R71 antibody is as shown in SEQ ID NO:3, and the heavy chain sequence of the R71 antibody is as shown in SEQ ID NO:4. It is also disclosed that the ratio of the R92 antibody to the R71 antibody in the combined monoclonal antibody preparation against rabies virus is (1-5):(1-5). The combined monoclonal antibody preparation against rabies virus of the present invention has a high affinity for rabies virus glycoprotein, has good neutralization ability and thermal stability, has good neutralization ability against a variety of street strains, and the two antibodies target different epitopes, providing a wider range of protection.

A method and kit are provided for detecting anti-drug antibodies (ADAs) against self-assembling trimeric biologics, such as XPro1595, a pegylated variant of soluble human Tumor Necrosis Factor, in biological samples, particularly human serum. The method employs an Affinity Capture Elution-Protein A / G (ACE-AG) assay that overcomes non-specific reagent interactions inherent in conventional bridging and standard ACE assays due to the biologic's dynamic monomer exchange. The assay utilizes a two-plate system: a streptavidin-coated plate with biotinylated trimeric biologic captures ADAs, which are eluted, neutralized, and transferred to a protein A / G-coated plate for specific immunoglobulin capture and detection with a sulfo-tagged trimeric biologic via electrochemiluminescence. The kit includes pre-coated plates, labeled biologics, elution and neutralization solutions, assay buffer, confirmatory reagent, and instructions. The assay offers a robust, specific, and sensitive solution for immunogenicity testing of trimeric biologics.

The invention relates to the technical field of antibodies, in particular to an anti-IL-17A nano antibody as well as a preparation method and application thereof. The invention provides a brand-new anti-IL-17A nano antibody, and the anti-IL-17A nano antibody shows relatively high affinity and neutralizing capacity of IL-17A.

The application discloses a high-expression-based CRM197 protein specific neutralization detection method and belongs to the technical field of biological detection. The method comprises the following steps: S1, strain resuspension: Corynebacterium diphtheriae is resuspended to obtain a bacterial suspension; S2, bacterial liquid culture: the bacterial suspension is inoculated into an iron trichloride YC liquid culture medium to be cultured to obtain a bacterial liquid; and S3, specific neutralization: the bacterial liquid is inoculated on Elek's medium to be subjected to virulence determination. The application establishes a novel CRM197 protein specific neutralization detection method, greatly improves the technical requirements of detection effect and time limit, and provides a novel detection method for biological science.

The present invention relates to specific monoclonal antibodies and their applications. The amino acid sequence of the light chain CDR1 of the monoclonal antibody is shown in SEQ ID No. 1, the amino acid sequence of CDR2 is shown in SEQ ID No. 2, and the amino acid sequence of CDR3 is shown in SEQ ID No. 3; the amino acid sequence of the heavy chain CDR1 of the monoclonal antibody is shown in SEQ ID No. 4, the amino acid sequence of CDR2 is shown in SEQ ID No. 5, and the amino acid sequence of CDR3 is shown in SEQ ID No. 6. The present invention uses recombinant monkeypox virus surface protein E8L as an antigen. The screened monoclonal antibodies have good affinity for the recombinant monkeypox virus surface protein E8L at the molecular and cellular levels, can significantly inhibit the infection efficiency of VACV replication-deficient strains in VERO cells, and have good in vitro neutralization effect against monkeypox virus.

This invention provides a human antibody with neutralizing ability against the JN.1 variant of SARS-CoV-2 and its application, belonging to the field of biomedical technology. The antibody comprises a heavy chain sequence as shown in SEQ ID NO:1 and a light chain sequence as shown in SEQ ID NO:2; or comprises a heavy chain sequence as shown in SEQ ID NO:3 and a light chain sequence as shown in SEQ ID NO:4. The antibody of this invention binds to the extracellular domain of the JN.1 variant spike protein via ECG. 50 The value was significantly lower than that of its parent antibody, with a binding capacity increase of 3 to 5 times or more. In the pseudovirus neutralization experiment, the antibody showed a half-maximal neutralizing concentration (IC50) against JN.1 pseudovirus. 50 The antibody also showed significantly better performance than the parent antibody, with a neutralizing activity increase of 3 to 24 times or more. Furthermore, the antibody's melting temperature exceeded 75°C, demonstrating good thermal stability.

The invention discloses a neutralizing swine monoclonal antibody pO18-40 and a neutralizing swine monoclonal antibody pO18-43 for foot and mouth disease virus type O. The amino acid sequences of a heavy chain variable region (VH) and a light chain variable region (VL) of the antibody pO18-40 are respectively as shown in SEQ ID No. 1 and SEQ ID No. 2; the amino acid sequences of VH and VL of the antibody pO18-43 are respectively as shown in SEQ ID No. 3 and SEQ ID No. 4. The antibody obtained by the invention is a full-swine-source antibody, can specifically neutralize the classical strain of the O-type foot-and-mouth disease virus, and can clearly distinguish the classical strain of the O / Cathay topological type from the variant strain of the O / Cathay topological type. A key antigen epitope recognized by the antibody is located at the 149th amino acid of a VP1 protein G-H ring, and the site is a key site of O / Cathay strain antigen variation and vaccine immune protection. The antibody provided by the invention provides an important tool and theoretical basis for serological detection of O-type FMDV, vaccine immune effect evaluation and optimal design of broad-spectrum vaccines.

A single-domain antibody (sdAb) which binds to head domain of VP6 derived from a human rotavirus A and has neutralization activity against different the rotavirus A types.

The application discloses camel-derived nanobodies of feline calicivirus and application thereof, and belongs to the field of biological medicine. The application takes feline calicivirus as an immunogen, and through steps of immunizing a camel, constructing a library, enriching sequencing and the like, two nanobodies VHH-V14 and VHH-V29 are screened, and the amino acid sequences of the two nanobodies are shown as SEQ ID NO. 1-2. Experimental results show that the nanobodies VHH-V14 and VHH-V29 are nanobodies with outstanding neutralization activity to feline calicivirus, and the IC 50 of the two nanobodies is 0.6536 mg / mL and 0.6931 mg / mL respectively, and the two nanobodies have good inhibiting effect on feline calicivirus. The application provides new biological materials for prevention and treatment of feline calicivirus infection, and provides new design theory and technical support for antibody treatment strategy of feline calicivirus, and has outstanding significance for clinical treatment of feline calicivirus infection.

The application discloses a recombinant nanobody against swine influenza virus of H1N1 subtype, and a preparation method and application thereof, and belongs to the field of biological medicines. A VHH sequence of the nanobody against swine influenza virus of H1N1 subtype is screened from a nanobody bacterial library of an immunized llama, the VHH sequence is directionally cloned into a pPIC9K vector to construct a recombinant eukaryotic expression plasmid, and the recombinant eukaryotic expression plasmid is transformed into a Pichia pastoris expression system to screen an expression engineering strain, and the recombinant nanobody is obtained after induction culture of the expression engineering strain. It is proved by in-vitro and in-vivo experiments that the recombinant nanobody developed in the application can neutralize CA / 04 and has a remarkable therapeutic effect. Therefore, the neutralizing nanobody provided in the application has considerable potential in treating CA / 04. The findings will provide experimental data for developing nanobody preparations against swine influenza virus of H1 subtype and provide a new strategy for preventing and controlling swine influenza.

The present invention provides neutralizing monoclonal antibodies targeting Nipah virus G protein and their uses, wherein the monoclonal antibodies can recognize Nipah virus G protein. The present invention uses NiV G protein as an antigen target, displays antigens on a ferritin nanoparticle platform to immunize mice, and screens out three monoclonal antibodies that can specifically bind to NiV G protein. Antibody epitope competition experiments found that the S1E2 and SB10 monoclonal antibodies among these three antibodies recognize new epitopes of NiV G protein that have not been reported before. In vitro neutralization experiments have demonstrated that these three antibodies have high in vitro neutralizing activity, and can neutralize both NiV-M and NiV-B strains, with the characteristics of high expression and good stability, and can be used to prepare virus detection products such as Nipah and Hendra or drugs for preventing and treating Nipah and Hendra virus diseases.

The invention provides anti-Ara h 2 antibodies (e.g., an anti-Ara h 2 neutralizing antibody) and methods of using the same, e.g., for treating and / or preventing peanut allergy or sensitivity. Also provided herein are anti-Ara h 2 antibodies and methods of using the same, e.g., for diagnostics and methods of monitoring peanut oral immunotherapy.

The invention relates to the technical field of biology, and aims to provide an M13 recombinant bacteriophage with PRRSV broad-spectrum neutralizing activity for displaying NPC and application of the M13 recombinant bacteriophage. A nano antibody-CD163 peptide fragment conjugate NPC shown as SEQ ID NO: 1 is displayed on the surface of the N end of capsid protein III of the M13 recombinant phage, 1-3 NPC molecules are contained on the surface of each phage particle, and the phage is used for neutralizing PRRSV (Porcine Reproductive and Respiratory Syndrome Virus). The recombinant bacteriophage provided by the invention is easy to prepare, extremely high in virus titer and excellent in thermal and pH stability; in Marc-145 and PAMs cells, the compound shows strong neutralizing activity on PRRSV II type 1, 3, 5 and 8 pedigree, so that the compound has broad-spectrum neutralizing capacity; the compound can be used as a PRRSV prevention and control preparation, and a new thought is provided for development of safe and efficient novel antiviral drugs or biological preparations.

The invention discloses a method for detecting an AAV neutralizing antibody. The method comprises the following steps: preparing a recombinant AAV vector AAV-Gluc carrying a Gaussian luciferase reporter gene; the method comprises the following steps: diluting AAV-Gluc according to a certain dilution step to prepare an AAV-Gluc working solution; diluting the sample to be detected; carrying out cell planking, culturing the target cells to form a uniform cell suspension, and adding the cell suspension into a cell culture plate; mixing the sample diluent with the AAV-Gluc diluent, and carrying out incubation; adding the incubated sample mixed solution into the cell culture plate, and carrying out virus infection and culture; obtaining a cultured cell supernatant, adding a detection solution, and detecting signal intensity; and analyzing and sorting the data to obtain the neutralization titer. The secreting type Gaussian luciferase reporter vector is used, target protein is secreted outside cells after being expressed, and detection is facilitated; the infected supernatant is directly sucked for detection, and the operation is convenient and fast; cell lysis and extraction are not needed, protein loss is reduced, and detection accuracy and precision are improved.

The sampling equipment comprises an adapter base, connecting bases are connected to the two ends of the adapter base, a neutralizing pipe is connected to one end of the adapter base, a neutralizing assembly is arranged in the neutralizing pipe, the neutralizing assembly comprises a rotating disc rotationally clamped in the neutralizing pipe, a side block is connected to the outer side of the rotating disc, and a first sliding groove is formed in the outer side of the neutralizing pipe; the side blocks are slidably clamped in the first sliding grooves, and the bottom end of the neutralizing pipe is connected with a fixing pipe. The arranged neutralization assembly can neutralize antibiotics in a blood sample of a patient before formal sampling is started, the inhibition effect of emergency antibiotic injection on germs in the blood sample of the patient is reduced as much as possible by neutralizing the antibiotics in the blood of the patient, and when the device is used, the concentration of the antibiotics in the blood of the patient is detected firstly; then sampling liquid is guided into the flow guide assembly through the liquid inlet hole, the flow guide assembly can guide the liquid into the neutralizing pipe, and the antibiotics are neutralized or absorbed through the neutralizing agent coated on the side wall of the connecting hole.

The invention relates to construction of a fusion antibacterial peptide expression vector as well as an expression purification method and a purification device thereof, and belongs to the technical field of bioengineering. The preparation method comprises the following steps: fusing an antibacterial peptide to a C terminal of RNase denaturation form Onconase (ONC) protein from Rana pipiens, constructing an expression plasmid, and transforming the expression plasmid into escherichia coli to obtain gt; the yield of the inclusion body form fusion peptide is 300 mg / L. Gt is realized through operations of inclusion body washing, redissolving, acid hydrolysis, neutralization and the like; and the low-cost large-scale production of the 95% high-purity antibacterial peptide is realized. As a carrier protein, ONC has the advantages of high expression quantity, low toxicity, small molecular weight, specific pH solubility and the like. In the purification process, the inclusion body is dissolved by using a urea solution, and the purity and the yield of the antibacterial peptide are further improved by combining cation exchange chromatography and an ultrafiltration technology. According to the method and the device, the purification efficiency is remarkably improved, the labor investment is reduced, and the method and the device are suitable for wide industrial application.

This invention belongs to the field of genetic engineering. By introducing a TC tag between amino acids 199 and 200 of the wild-type bluetongue virus non-structural protein NS2, a recombinant virus was rescued. Analysis of viral plaques and growth curves revealed that the recombinant virus formed plaques of similar size to the wild-type virus, and their growth curves showed no significant difference. Immunofluorescence and FlasH-EDT2 staining of NS2 showed that the fluorescence of NS2 labeled with anti-NS2 antibody overlapped with that labeled with FlasH-EDT2. Three-dimensional imaging of FlasH-EDT2-labeled NS2 revealed that the viral inclusion bodies exhibited fluorescence around the periphery but no fluorescence inside, displaying an overall "core-shell" hierarchical structure. This recombinant bluetongue virus can be used for tracing and localizing NS2 protein and viral inclusion bodies in living cells, and can also be applied to BTV neutralization assays, antiviral drug screening, and other related research.

The present application relates to the development and application of a kind of nanobody 2A5 for neutralizing broad-spectrum respiratory syncytial virus, and specifically relates to a kind of nanobody, which includes CDR1 as shown in SEQ ID NO:3, CDR2 as shown in SEQ ID NO:4, and CDR3 as shown in SEQ ID NO:5.The nanobody provided by the present application has the characteristics of high affinity and high neutralization, can effectively neutralize respiratory syncytial virus A2 and B strain, and the neutralization capacity is better than that of marketed antibody Nirsevimab.Therefore, the nanobody has great potential for further development as a candidate for preventing and treating respiratory syncytial virus, and also provides a scientific basis for the development of other antiviral therapies for respiratory syncytial virus.

The invention relates to the field of chemical analysis and detection, in particular to a method for rapidly detecting the concentration of industrial concentrated acid based on neutralization reaction, which comprises the following steps: 1, theoretical calculation; converting molar concentration according to acid density and mass fraction; 2, accurately measuring the volume of target acid by using a pipette; 2, injecting 100-150ml of pure water into the triangular flask, and adding 3-5 drops of phenolphthalein; 3, slowly injecting an acid sample into the triangular flask, and slightly shaking and uniformly mixing; 4, adding 10.00 ml of NaOH solution by using the pipette, and oscillating for 10-20 seconds; 3, result interpretation: transparency and colorless: acid is excessive when the acid concentration reaches the standard, pH is less than or equal to 8.2 after neutralization, pink lasts for 20-30 seconds, and reinspection is needed when the acid concentration is insufficient and the pH is greater than or equal to 8.3. The method has the beneficial effects that 1, a reverse judgment method is adopted, namely the conventional end-point titration is replaced by fixed alkali amount, so that the detection time is shortened to 2 minutes; 2, safety design: pre-dilution reduces the risk of acid mist, and only a trace amount of raw acid is needed to avoid acid corrosion; 3, multi-acid adaptation: compatibility with detection of different types of concentrated acids is realized through pipetting volume adjustment;

The invention relates to the technical field of protein detection, in particular to a detection method and a kit for detecting an anti-Nephrin antibody. The detection method comprises the following steps: capturing an anti-Nephrin antibody in a sample to be detected on an SA matrix by adopting biotinylated Nephrin protein to obtain a first compound; carrying out acidolysis desorption and neutralization on the first compound, and directionally fixing an antibody by Protein G to obtain a second compound; aiming at the second compound, detecting by adopting a biotin-streptavidin-HRP (Horse Radical Polymorphism) system; the acidolysis desorption comprises the following steps: treating the first compound for 10-20 minutes by adopting an acid solution with the pH value of 2-3 under the conditions that the temperature is 35-40 DEG C and the speed is 500-800 rpm. The method provided by the invention can accurately and quickly detect the level of the anti-Nephrin antibody in the sample, and has important application value.

The present invention discloses a neutralizing anti-idiotypic antibody of an anti-D antibody, comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises at least one of HCDR1, HCDR2, and HCDR3 sequences, wherein: the amino acid sequence of the HCDR1 is shown in SEQ ID No. 1; the amino acid sequence of the HCDR2 is shown in SEQ ID No. 2; and the amino acid sequence of the HCDR3 is shown in SEQ ID No. 3. This neutralizing anti-idiotypic antibody has the ability to neutralize anti-D-mediated hemagglutination of red blood cells and can be used to detect mixed samples containing multiple blood type antibodies unexpectedly containing anti-D. It can also be used for risk assessment and clinical diagnosis and treatment of Rh-HDFN.