30 results about "Hemagglutination" patented technology

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.

The invention belongs to the field of biology, and discloses preparation and application of a non-methanol-induced expression H5 subtype avian influenza virus subunit antigen and a vaccine thereof, PDAK is applied to pichia pastoris expression of avian influenza virus H5 subtype HA protein, an HA protein multiple-insertion recombinant strain with a PDAK promoter is successfully constructed, and in the research process, it is found that the H5 subtype avian influenza virus subunit antigen and the vaccine thereof have the advantages that the H5 subtype avian influenza virus subunit antigen and the vaccine thereof can be used for preparing the H5 subtype avian influenza virus subunit antigen; by adopting the recombinant plasmid disclosed by the invention, multiple insertions can be generated in a pichia pastoris strain, the target protein expression quantity can be obviously improved, and the foreign protein expression efficiency of the recombinant plasmid is superior to that of a PAOX1 promoter and a PGAP promoter; according to the invention, the HA protein of the PDAK promoter is multiply inserted into the recombinant strain to express the HA protein to prepare the subunit vaccine, after the vaccine is immunized, a higher hemagglutination inhibition antibody level can be generated aiming at homologous strains, and the survival rate of experimental chickens after challenge is 100%.

This invention discloses a method for detecting antigen content, comprising the following steps: diluting the test sample and a reference sample to specific concentrations; serially diluting the test sample and reference sample to specific concentrations using a diluent on a 96-well hemagglutination plate; adding sensitized red blood cell suspension to wells of different dilutions of the test sample and reference sample, mixing, allowing to stand, and then determining the agglutination results of each well; scoring and recording the agglutination results of each well; summing the scores of each test sample and recording it as A, and summing the total scores of each reference sample and recording it as B; calculating the antigen content of the test sample according to the formula: antigen content of the reference sample × 2. (A‑B) / 16 This method is convenient and fast. The test results are calculated using the self-developed method and compared with the test results of the reference sample, which can obtain relatively accurate test results and solve the problem of inconsistent test results of different batches of sensitized red blood cell suspensions.

The invention discloses a recombinant nano antibody of a waterfowl H9N2 avian influenza virus, a recombinant expression vector, a recombinant strain and application thereof, and relates to the field of biology. The amino acid sequence of the recombinant nano antibody is SEQ ID NO: 2, and the nucleotide sequence of the recombinant nano antibody is SEQ ID NO: 1. The invention provides a recombinant expression vector containing the coding gene of the recombinant nano-antibody and a recombinant strain for expressing the recombinant nano-antibody. The recombinant nano antibody is applied to preparation of a waterfowl H9N2 avian influenza resisting medicine or a waterfowl H9N2 avian influenza diagnostic reagent. The yield of the recombinant nano antibody reaches 1.5 g / L, the hemagglutination inhibition activity titer for resisting the waterfowl H9N2 avian influenza virus is 4 log2, and the recombinant nano antibody has remarkable biological activity and can be used for preparing a novel nano antibody preparation for preventing and / or treating the waterfowl H9N2 avian influenza.

Provided herein are anti-CD47 antibodies and immunologically active fragments thereof that have prolonged antitumor effects and cause low or no level of red blood cell or platelet depletion and hemagglutination. Also provided are as well as pharmaceutical compositions containing such antibodies or antibody fragments, as well as methods of treatment using such antibodies, e.g., as single agents or in combination with other therapeutic agent(s).

The present invention belongs to the field of interferon genetic engineering, and discloses recombinant pigeon alpha interferon and its application in the preparation of medicines for resisting pigeon paramyxovirus infection. The present invention constructs recombinant pigeon interferon piINF 5, realizes efficient expression and purification in CHO cell expression system, significantly prolongs its half-life and improves stability. Recombinant pigeon interferon piINF 5 has no toxic effect on DF1 and VERO cells, and can effectively stimulate the transcription level of interferon-stimulated genes, can significantly inhibit the replication of pigeon paramyxovirus, reduce viral copy number and hemagglutination value. In vivo animal attack experiment shows that the recombinant pigeon interferon piINF 5 can well protect pigeons from the attack of pigeon paramyxovirus, and the survival rate reaches 80%, and there is no detoxification in the first five days after the attack, and only 20% of animals have detoxification in the later stage. The present invention provides an efficient and stable new drug selection for the treatment of pigeon paramyxovirus disease, with good commercial application prospects.

The invention relates to the technical field of biology, and discloses an influenza A virus H5 protein mutant and application thereof. In order to solve the problems of weak cross immune response to epidemic strains, insufficient stability of wild type HA protein, low immunogenicity and the like of the existing H5N1 vaccine, a stable soluble H5 protein mutant is constructed through a sequence modification strategy. Specifically, the 332th to 380th amino acids in the HA protein of a wild type influenza A virus H5N1 are replaced by SPGCAT, and point mutations T386C, L418P and H456P are introduced to obtain the mutant. The mutant can significantly improve the structural stability of trimer protein, improve the expression level of recombinant protein, broaden the cross protection spectrum, enhance the level of immunogenic hemagglutination inhibition antibodies and improve immune response balance.

The present invention describes hemagglutination inhibition (HAI) assay methods to determine candidate subunit protein vaccines in the absence of using source viruses.

The present invention belongs to the technical field of biomedicine, and specifically relates to a recombinant influenza virus protein, a vaccine, and a preparation method therefor and the use thereof. To overcome the defects of existing influenza vaccines with respect to safety and high toxic side effects, a recombinant influenza virus protein is provided, which has an amino acid sequence as shown in at least one of SEQ ID No. 1 to SEQ ID No. 6. Recombinant hemagglutinin proteins of H1N1 and H3N2 subtypes, and Victoria and Yamagata lineages are expressed and purified by means of an insect baculovirus expression system, and are mixed with an adjuvant to prepare a highly immunogenic recombinant bivalent, trivalent or quadrivalent recombinant influenza virus protein vaccine. The prepared vaccine can induce high levels of specific IgG antibodies and hemagglutination-inhibiting neutralizing antibodies, the addition of adjuvant WGa01 improves the immunogenicity of the vaccine, and the provided vaccine is suitable for people allergic to eggs and has high safety.

The invention belongs to the field of interferon gene engineering, and discloses a recombinant pigeon interferon alpha and application thereof in preparation of a medicine for resisting pigeon paramyxovirus infection. The recombinant pigeon interferon piINF-5 is constructed, efficient expression and purification in a CHO cell expression system are achieved, the half-life period of the pigeon interferon piINF-5 is remarkably prolonged, and the stability of the pigeon interferon piINF-5 is improved. The recombinant pigeon interferon piINF-5 has no toxic effect on DF1 and VERO cells, can effectively stimulate the transcriptional level of interferon stimulating genes, can significantly inhibit the replication of pigeon paramyxovirus, and reduces the virus copy number and hemagglutination price. Animal in-vivo challenge experiments show that the recombinant pigeon interferon piINF-5 can well protect pigeons from attacking pigeon paramyxoviruses, the survival rate reaches 80%, no toxin is expelled in the first five days after challenge, and only 20% of animals are expelled in the later period. The invention provides an efficient and stable novel drug choice for treatment of the pigeon paramyxovirus disease, and has a good commercial application prospect.

The invention discloses an anti-influenza A virus micropeptide uYIPF2 coded by an upstream open reading frame of a YIPF2 gene and application thereof, and relates to the technical field of biological medicines. The micropeptide uYIPF2 is coded by an upstream open reading frame (uORF) of a YIPF2 gene, contains 87 amino acids, and has a specific sequence as shown in SEQ ID No.1. A hemagglutination test and a plaque test prove that the micropeptide uYIPF2 can significantly reduce the HA titer and the virus load of the influenza A virus, and can significantly inhibit the replication of the influenza A virus. The invention reveals that the uYIPF2 micro-peptide has an anti-influenza A virus biological function for the first time, and provides application of the uYIPF2 micro-peptide in preparation of drugs for preventing and treating influenza A virus infection. The invention provides a brand-new drug target and a treatment strategy for the field of flu prevention and treatment.

An adenovirus that reduces the extent to which neutralizing antibodies bind to the adenovirus has an AB loop that includes a sufficient portion of the amino acid sequence VTINRSA (amino acids 8-14 of SEQ ID NO:2), TYMLSRN (amino acids 8-14 of SEQ ID NO:3), or STMGTSH motif (amino acids 8-14 of SEQ ID NO:4) to reduce binding of neutralizing anti-adenovirus antibodies compared to an adenovirus having a wild-type AB loop. In some cases, another aspect, the adenovirus has an AB loop that includes a sufficient portion of the amino acid sequence VTINRSA (amino acids 8-14 of SEQ ID NO:2), TYMLSRN (amino acids 8-14 of SEQ ID NO:3), or STMGTSH motif (amino acids 8-14 of SEQ ID NO:4) to reduce hemagglutination compared to an adenovirus having a wild-type AB loop. The adenovirus can be used to deliver therapy to a target cell to which the AB loop binds.

The invention discloses a histophilic somniphilus, a hemagglutination antigen, a hemagglutination inhibition antibody detection method, a kit and application of the histophilic somniphilus and the hemagglutination antigen. The method comprises the following steps: screening a histophilic sleep bacteria Yunnan isolate ZY25055 with high hemagglutination activity, culturing, and carrying out beta-propiolactone inactivation to prepare the histophilic sleep bacteria hemagglutination antigen. 1% hydroformylated sheep red blood cells are prepared by collecting sheep blood. Immunizing goat posterior jugular vein blood sampling with beta-propiolactone inactivated histophilic sleep bacteria liquid for three times, and centrifugally separating serum to prepare histophilic sleep bacteria standard positive serum. The method and the kit for detecting the histophilic sleeping bacteria hemagglutination inhibition antibody are established by utilizing the prepared histophilic sleeping bacteria hemagglutination antigen, 1% hydroformylated sheep red blood cells and histophilic sleeping bacteria standard positive serum, the method has no cross reaction with positive serum of common pathogens of cattle and sheep, and the method has the advantages of strong specificity, simplicity and convenience in operation, no need of complex equipment and the like, and can be used for detecting the histophilic sleeping bacteria hemagglutination inhibition antibody. The kit can be used for detecting the histophilic somniphilus antibody in bovine serum, sheep serum and goat serum.

The present invention describes hemagglutination inhibition (HAI) assay methods to determine candidate subunit protein vaccines in the absence of using source viruses.

The invention provides a method for preparing a Newcastle disease virus antigen by a cell suspension culture process, and relates to the technical field of virus antigen and vaccine preparation. The suspension culture method comprises the following steps: inoculating full-suspension Newcastle disease virus production cells into an F807k basal culture medium according to a certain density for cell culture, adding an NF01d supplementary culture medium after the cell culture is performed for 70-74 hours, adding an F808Vb supplementary culture medium after the cell culture is performed for 94-98 hours, inoculating Newcastle disease virus according to a virus inoculation amount of 0.1-0.5%, and culturing for 20-30 hours. And 38-42 hours after virus inoculation, adding the NF01d supplementary culture medium again, and expressing the Newcastle disease virus antigen at 28-38 DEG C. According to the optimized full-suspension culture process, higher antigen expression quantity can be obtained, the virus yield is improved, the hemagglutination titer of the obtained Newcastle disease virus antigen reaches 12log2 for the first time, and the protective efficacy is improved.

The present invention discloses a method for detecting the hemagglutination titer of an influenza virus antigen sample, comprising the following steps: S1. Lysis treatment: adding a lysing agent to the influenza virus antigen sample and the influenza virus antigen hemagglutination titer reference product respectively for lysis to obtain a lysed influenza virus antigen sample and a lysed influenza virus antigen hemagglutination titer reference product; the influenza virus antigen sample is an intermediate produced in each process during the production of the monovalent bulk of the influenza virus split vaccine; S2. Hemagglutination titer detection: detecting the hemagglutination titer of the lysed influenza virus antigen sample and the hemagglutination titer of the lysed influenza virus antigen hemagglutination titer reference product respectively; S3. Calculating the standard value of the hemagglutination titer of the influenza virus antigen sample. The detection method of the present application has good precision, good repeatability and good accuracy, and can be used for the quality control evaluation of intermediates in each process during the production of the monovalent bulk of the influenza virus split vaccine.

The present invention relates to the field of biological vaccine research and development technology, and in particular, to a VSV vector-based highly pathogenic H5 subtype avian influenza inactivated vaccine and a method for preparing same. The method comprises: using a vesicular stomatitis virus with the deletion of the surface envelope protein gene as a vector, inserting the NA gene and the HA gene of an H5 subtype highly pathogenic avian influenza virus strain between the M gene and the L gene of the vector in the form of an expression cassette, and acquiring a recombinant virus by means of rescue via reverse genetic technology; inoculating the recombinant virus into susceptible cells, culturing for replication and proliferation, and then harvesting a viral solution; inactivating the viral solution to acquire an inactivated viral solution; and adding the inactivated viral solution to a pharmaceutically or veterinarily acceptable carrier, excipient, medium, or adjuvant to acquire an inactivated vaccine. The provided H5 subtype avian influenza inactivated vaccine can induce a high hemagglutination inhibition titer of avian influenza virus on day 21 after chick immunization. The provided inactivated vaccine has good safety and immunogenicity.

The invention discloses a recombinant nano antibody for resisting H9 subtype avian influenza virus HA protein and application of the recombinant nano antibody, and relates to the field of bioengineering, and the amino acid sequence of the recombinant nano antibody is SEQ ID NO: 2; the nucleotide sequence for coding the amino acid sequence of the recombinant nano antibody is SEQ ID NO: 1. The yield of the recombinant nano-antibody is 1.84 mg / mL, the recombinant nano-antibody is only specifically combined with H9 virus and HA protein, has hemagglutination inhibition activity and HI titer of 8.3 log2, is the first nano-antibody with hemagglutination inhibition activity reported at present, has half effective inhibition concentration IC50 of 16.31 mu g / mL, can significantly reduce the detoxification level of the virus in chicks, has high biological activity, and can be used for preparing the recombinant nano-antibody. The strain can be applied to prevention and / or treatment of H9 subtype avian influenza, provides a novel, efficient and customizable biological preparation platform for prevention and control of avian influenza, and has important scientific research value and industrialization prospect.

The present invention discloses a monoclonal antibody that broadly binds to the HA protein of the multi-branched H5Nx subtype avian influenza virus and its application. The monoclonal antibody is named 1G10, belongs to the IgG1 subclass, comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises amino acid sequences as shown in SEQ ID NO.5-7. H CDR1, V H CDR2 and V H CDR3, the light chain variable region comprises amino acid sequences as shown in SEQ ID NO.8-10 L CDR1, V L CDR2 and V L The monoclonal antibody specifically recognized the HA protein in indirect immunofluorescence assays, protein immunoblotting, and neutralization tests. Furthermore, the monoclonal antibody was able to undergo specific hemagglutination inhibition with H5Nx subtype avian influenza viruses, including H5N1, H5N6, and H5N8, from different HA gene evolutionary branches, such as 2.3.2.1d, 2.3.4.4b, and 2.3.4.4d, indicating that the antigenic epitope recognized by the 1G10 monoclonal antibody is a broad-spectrum neutralizing epitope for H5 subtype avian influenza viruses. Chicken embryo immune escape assays and sequence alignment revealed that the neutralizing epitope recognized by the 1G10 monoclonal antibody includes amino acid positions 170 and 231 of the HA protein, and that mutations significantly affect antigenicity.

Provided are an antibody or an antigen-binding fragment thereof that specifically recognizes CD47 and methods of making the same and using the same. The antibodies provided herein exhibit both induction of phagocytosis of cancer cells and have very low to no hemagglutination and phagocytosis of RBCs, showing improved efficacy to toxicity ratio for cancer treatment.

The invention belongs to the technical field of biological medicines, and particularly relates to a recombinant influenza virus protein, a vaccine and a preparation method and application thereof. In order to overcome the defects of high safety and toxic and side effects of the existing influenza vaccine, the invention provides a recombinant influenza virus protein, the amino acid sequence of which is as shown in at least one of SEQ ID No.1-SEQ ID No.6. According to the invention, H1N1 subtype, H3N2 subtype, Victoria line and Yamagata line recombinant hemagglutinin proteins are expressed and purified through an insect baculovirus expression system, and are mixed with an adjuvant to prepare a recombinant divalent, trivalent or tetravalent recombinant influenza virus protein vaccine with high immunogenicity. The prepared vaccine can induce high-level specific IgG antibodies and hemagglutination inhibition neutralizing antibodies, the immunogenicity of the vaccine is improved by adding the adjuvant WGa01, and the vaccine is suitable for egg allergic people and high in safety.

The present invention relates to a chimera formed of peptide epithedes derived from hemagglutinin and neuraminidase characterized in that it consists of SEQ ID NO.4.

The invention discloses a chimeric monoclonal antibody specifically combined with feline parvovirus and application of the chimeric monoclonal antibody, and belongs to the technical field of antibody engineering. The chimeric monoclonal antibody is formed by chimeric combination of a mouse source monoclonal antibody variable region and a cat source antibody constant region; the variable region of the mouse monoclonal antibody comprises a heavy chain variable region with a sequence as shown in SEQ ID NO.3 and a light chain variable region with a sequence as shown in SEQ ID NO.4. The hemagglutination inhibition titer of the chimeric monoclonal antibody against FPV reaches 1: 4194304, IC50 is 0.0008 mg / mL, and the chimeric monoclonal antibody can completely protect an FPV infected cat model and has broad-spectrum neutralizing activity on various parvovirus variants. The invention fills the blank of the current specific therapeutic drug for feline panleucopenia, and has the advantages of high efficiency, low immunogenicity and large-scale production.

The utility model provides an avian influenza hemagglutination inhibition test analyzer, which comprises a rack, and further comprises a carrier, a rotating assembly, a circulating assembly, a light source plate and a mounting plate, and a plurality of carrier holes are formed in the carrier and are used for placing hemagglutination plates to be detected. By means of the design, the device has more advantages in batch testing, and the working efficiency is improved. According to the design, the accuracy and the efficiency of an avian influenza hemagglutination inhibition test are improved, the time and the labor cost of a laboratory can be saved, the manual operation is reduced, the automation degree of the test is improved, the test efficiency is improved, the personal error is reduced, and a shot sample result can be recorded; and the work of scientific research and clinical detection is more efficient and quicker. Through reasonable structural design and intelligent function configuration, an efficient, accurate and reliable solution is provided for related research and clinical detection, and the requirements of modern laboratories for diversification and high efficiency of equipment are met.

The invention relates to a CD47 antibody and application thereof, in particular to a monoclonal antibody for recognizing CD47, more particularly to a CD47 antibody which does not cause significant levels of red blood cell agglutination, red blood cell reduction and thrombocytopenia, and relates to a method for preparing the antibodies and application of the monoclonal antibody in preparation of medicines for treating cancers or infections.

1. Name of the Design Product: Hemagglutination Analyzer (Medical Examination). 2. Use of the Design Product: For blood testing. 3. Design Key Points of the Design Product: Lies in the shape. 4. Picture or Photograph that Best Illustrates the Design Key Points: Perspective View 1.

The invention relates to the technical field of hemagglutination inhibition antigen preparation, in particular to a preparation method of a formaldehyde-inactivated A-type avibacterium paragallinarum hemagglutination inhibition antigen. According to the technical scheme, the method comprises the following steps: preparing an antigen repair liquid, wherein the antigen repair liquid comprises ethylenediamine tetraacetic acid, sodium citrate dehydrate, tween-20, triethanolamine and purified water; according to the invention, the hemagglutination activity of formaldehyde inactivated A type avibacterium paragallinarum is successfully recovered through an optimized antigen repair liquid and a mild repair process, and the problem of titer loss caused by antigen epitope shielding in a traditional inactivation process is solved; according to the method, the inactivated bacterial liquid is directly utilized, complex culture and crushing steps are omitted, the process is remarkably simplified, the cost is reduced, and meanwhile, the antigen stability and the preservation period are improved; the kit is simple in structure, convenient and safe to operate and high in antigen specificity, can be accurately used for serum antibody detection and vaccine effect evaluation, and provides efficient and reliable technical support for diagnosis, prevention and control of avian infectious rhinitis.

The invention discloses an anti-H7N9 influenza virus monoclonal antibody and an application thereof. The anti-H7N9 influenza virus monoclonal antibody disclosed by the invention has relatively high affinity with various H7N9 viruses, the affinity with H7N9-AH13HA is at a pM level, and the affinity with H7N9-SF003HA and H7N9-SD12HA is at an nM level; the neutralizing activity (IC50) on H7N9 / AH13 and H7N1 / SF003 can reach 0.01 mu g / mL, and the neutralizing activity on H7N9 / AH13 and H7N1 / SF003 can reach 0.005 mu g / mL; the hemagglutination titer to H7N9 / AH13 is 0.97 mu g / mL, the hemagglutination titer to H7N1 / SF003 is 1.95 mu g / mL, and both the H7N9 / AH13 and the SF003 have very high neutralization titers; the kit can be used for detecting the H7N9 influenza virus and / or the HA protein of the H7N9 influenza virus, and preventing and / or treating diseases caused by the H7N9 influenza virus.

This invention relates to the field of bioengineering, and more particularly to sgRNA, MDCK cells, and their applications. This invention provides sgRNA having the nucleotide sequences shown in SEQ ID NO:1 to SEQ ID NO:4. This invention employs a CRISPR-Cas9 gene editing method, which, compared to existing RNAi methods, offers higher specificity and lower off-target risk. Compared to MDCK cells, PCR detection showed a reduction of over 90% in RNA levels in IRF7- / - cells, Western blotting showed a significant decrease in IRF7 protein expression, and viral hemagglutination assays showed higher titers for influenza virus compared to the original cells. Tumorigenicity assays showed that the gene knockout cells retained low tumorigenicity. Compared to existing technologies, the cells of this invention exhibit both low tumorigenicity and significantly increased viral yield, making them suitable for influenza vaccine production.

The invention discloses a novel red blood cell agglutination and red blood cell agglutination inhibition test detection method, which comprises the following steps of: after adding a to-be-detected sample into 1% of pig red blood cell suspension to react, sucking 50 microliters of supernatant liquid into a 96-hole ELISA (enzyme-linked immunosorbent assay) plate, and reading at the wavelength of 450 nm by using a microplate reader; calculating the average value of the OD values of the four multiple holes of each dilution degree of the sample to be detected, substituting the dilution times and the average value of the OD values into fitting software ELISACalc, and fitting by adopting a four-parameter fitting mode to obtain an S-shaped curve and an equation; the linear parameter R2 of the obtained S-shaped curve is observed, and when R2 is larger than or equal to 0.97, the linear parameter meets the requirement, and the test result is credible; and substituting the OD average value of the control sample into the equation to obtain a corresponding dilution multiple, namely the erythrocyte agglutination valence or erythrocyte agglutination inhibition valence of the sample to be detected. The scientificity and accuracy of the erythrocyte agglutination and erythrocyte agglutination inhibition test method are improved.