93 results about "Serum antibody" patented technology

The invention provides a characteristic protein of serotype I chicken Marek's disease virus and detection application, and belongs to the technical field of virus detection. The invention provides a recombinant gene derived from serum I type chicken Marek's disease virus. The nucleotide sequence of the recombinant gene is shown as SEQ ID No. 1. The recombinant gene disclosed by the invention is used for coding the MDV-1 recombinant PP38 protein, and the MDV-1 recombinant PP38 protein can be applied to enzyme-linked immuno sorbent assay (ELISA) detection. The invention also provides an ELISA detection kit constructed by using the MDV-1 recombinant PP38 protein, and the ELISA detection kit can be used for clinical MDV-1 serum antibody detection, SPF chicken screening, exogenous virus antibody detection of poultry live vaccines or virus seeds, epidemiological investigation and the like, so as to prevent and control transmission of MDV-1 infection.

The invention belongs to the field of antibody development, and discloses a screening method and application of a human-mouse protein high homologous target antibody based on a fully humanized antibody mouse. Aiming at the problem of weak antibody response caused by immune tolerance of human-mouse high homologous targets (protein homology is greater than or equal to 95%), the following scheme is provided: in embryonic stem cells (ES cells) of HUGO-Mabfully humanized antibody transgenic mice, a mouse target gene (such as ACVR2A) is knocked out through a Turbo Knockout technology, and homozygous knockout ES clones are screened; carrying out microinjection on the clones to the whitened B6 mouse blastocyst, and transplanting a pregnant mouse to obtain a Founder mouse; the Founder mouse is subjected to target antigen immunization for more than or equal to 4 times (the Freund's complete adjuvant is used for the first time), and the titer of the serum antibody is detected. According to the invention, 100% homozygous knockout chimeric efficiency is realized in the Founder stage, the mouse construction period is shortened from traditional 8-10 months to 3-4 months, and the diversity and affinity of the antibody are significantly improved (titer reaches 1: 729,000). The obtained antibody can be used for preparing medicines for treating tumors or autoimmune diseases.

The invention discloses an AIE fluorescence immunochromatography test strip for detecting a toxoplasma gondii antibody as well as a preparation method and application of the AIE fluorescence immunochromatography test strip, and belongs to the technical field of production of veterinary biological diagnosis products. The test strip provided by the invention comprises a bottom plate, a sample pad, a combination pad, a nitrocellulose membrane and a water absorption pad, wherein the sample pad, the combination pad, the nitrocellulose membrane and the water absorption pad are bonded on the bottom plate in an end-to-end connection manner; the combination pad is coated with AIE microsphere labeled protein; the nitrocellulose membrane contains a detection line T and a quality control line C; the test strip prepared by adopting a double-antigen sandwich antibody detection method can be used for detecting serum antibodies in animal bodies such as pigs, cats and the like. Compared with a colloidal gold test strip on the market, the colloidal gold test strip has the advantages that the sensitivity is improved by 32 times, the batch-to-batch difference is less than 5%, the test performed by adopting 40 parts of pig serum and 30 parts of cat serum shows that the specificity, the stability and the accuracy are excellent, the operation is simple, the cost is low, and an accurate result can be obtained only in 10 minutes; and a rapid and effective detection method is provided for epidemiological investigation and disease diagnosis.

The application discloses application of a cryptosporidium cgd6_2310 protein as an oocyst outer wall marker protein, an amino acid sequence of which is shown in the sequence table SEQ ID N0.1; a cryptosporidium oocyst wall protein specific polypeptide, an amino sequence of which is CLTPQSIERKEGTIID or ELDSSRTPVNETINC; an anti-cryptosporidium oocyst outer wall protein antibody, which can specifically bind to the outer surface of the cryptosporidium oocyst outer wall; an anti-cryptosporidium oocyst outer wall specific polypeptide antibody, which is an antibody prepared by using the cryptosporidium oocyst wall protein specific polypeptide; application of the anti-cryptosporidium oocyst outer wall protein antibody or the anti-cryptosporidium oocyst outer wall protein specific polypeptide antibody in detection of cryptosporidium; the polypeptide rabbit serum antibody has good sensitivity; in detection of samples in the environment, the polypeptide antibody only recognizes the cryptosporidium oocyst and does not cross react, and has good specificity.

The invention discloses a B cell epitope on the surface of glyceraldehyde-3-phosphate dehydrogenase protein, a nucleic acid molecule of the B cell epitope and application of the B cell epitope to detection of mycoplasma hyopneumoniae infection. The B cell epitope can be expressed on the surface of an inert carrier S9 thallus in a functional exhibition mode, an LN7-antibody direct-mediated agglutination test detection method established based on the LN7 expressed on the surface of the S9 thallus in the functional exhibition mode is adopted, the LN7 only specifically recognizes a mycoplasma hyopneumoniae infected serum antibody and a vaccine immune serum antibody containing a GAPDH antigen, a specific binding reaction is conducted, and the mycoplasma hyopneumoniae infected serum antibody and the vaccine immune serum antibody containing the GAPDH antigen can be detected. And cross reaction and non-specific binding reaction with other mycoplasma pathogens, porcine respiratory disease pathogen infection serum and vaccine immune serum are avoided. The diagnosis method is expected to become an important technical means for accurate diagnosis of mycoplasma hyopneumoniae infection and monitoring and evaluation of the immune effect of the mycoplasma hyopneumoniae vaccine.

The invention belongs to the technical field of food additives, and particularly relates to a nutrition enhancer as well as a preparation method and application thereof. The nutrition enhancer comprises a component I derived from plant polypeptide, a component II derived from iron, a component III derived from calcium, a component IV derived from zinc and an absorption enhancer, the absorption enhancer is composed of spermine and rhizoma atractylodis polysaccharide, wherein the spermine is loaded by silicon dioxide microspheres coated with modified chitosan. Under the action of the absorption enhancer, the nutrition enhancer disclosed by the invention can promote effective absorption of calcium, iron and zinc, and meanwhile, the content of serum antibodies can be increased.

The invention provides a preparation method of an H5 and H7 subtype recombinant avian influenza virus hemagglutination inhibition test antigen, which overcomes the defects that in the existing avian influenza virus antibody hemagglutination inhibition test, a standard substance cannot accurately detect the virulence of a strain to be detected and cannot truly reflect the immune effect of an avian influenza vaccine, and the H5 and H7 subtype recombinant avian influenza virus hemagglutination inhibition test antigen can be obtained by adjusting inactivation and purification processes. The pH value is adjusted, so that impure protein is separated out more easily, components influencing a detection result are reduced, the binding capacity of hemagglutinin is changed, the activity of the hemagglutinin is more easily bound with a serum antibody, the impure protein, inorganic salt and urate in an antigen are removed, and errors in the detection process are reduced. The method provided by the invention is high in specificity, the detection result is more accurate and reliable, and the effectiveness of the vaccine can be better evaluated.

The invention belongs to the technical field of bioengineering, and discloses a monoclonal antibody specifically combined with clostridium septicum alpha toxin recombinant protein, a hybridoma cell strain and application. The invention provides a monoclonal antibody which is specifically combined with clostridium septicum alpha toxin protein and a hybridoma cell strain clostridium septicum alpha toxin-8H2 which secretes the monoclonal antibody and has the preservation number of CCTCC NO: C202535, the monoclonal antibody only reacts with the clostridium septicum alpha toxin protein, and does not have cross reaction with other pathogens which infect cattle and sheep; the specificity is extremely high; according to the kit, a competitive ELISA technology is utilized, a mouse monoclonal antibody specifically combined with clostridium septicum alpha toxin recombinant protein is taken as a competitive antibody, serum antibodies of various animal sources can be detected, and species limitation of sample sources is avoided. Besides, the kit is good in repeatability, relatively loose in technical requirements during operation, capable of being widely popularized and used in production, and applied to clostridium septicum epidemiological investigation and vaccine immune level monitoring.

Provided are a Dabie bandavirus mRNA vaccine and a preparation method therefor. Specifically an optimized mRNA molecule is provided, and is cloned into a pGEM-3Zf(+)mRNA vaccine vector; the plasmid is linearized by means of enzyme digestion, and subjected to capping and poly(A) tail addition by means of an in-vitro transcriptase method to prepare an mRNA; the obtained mRNA is transfected into eukaryotic cells, and it is verified by means of an immunoblotting experiment that the mRNA can be expressed in vitro; and the mRNA is encapsulated by a lipid nanoparticle delivery system to obtain the mRNA vaccine, which, after immunizing mice, can induce the generation of a relatively high antibody level in serum. A viral neutralization test further proves that the immune serum can bind to viruses to prevent the viruses from infecting cells.

The invention discloses a diagnostic method based on mouse lyme disease serology indirect ELISA laboratory, and relates to the technical field of immunoserology detection.The diagnostic method comprises the steps that experimental conditions are optimized through a system, and the optimal reaction system is determined as follows: the antigen coating concentration is 0.2 microgram / microliter, the serum dilution ratio is 1: 200, and the enzyme-labeled secondary antibody concentration is 1: 2000. 0.8), and meanwhile, the antigen dosage is obviously reduced. Dynamic analysis of the antibody shows that the level of the antibody infecting mouse serum reaches a peak 12-25 days after inoculation, and the threshold value of the specific antibody ratio (ArB%) is 71.43%. Researches prove that the method has the characteristics of simplicity and convenience in operation, economy and high efficiency, and is suitable for primary laboratories and large-scale screening. The method provides reliable standardized technical support for lyme disease laboratory diagnosis and epidemiological monitoring.

The present invention discloses a B cell epitope on the surface of a glyceraldehyde-3-phosphate dehydrogenase protein, its nucleic acid molecule, and its application in detecting Mycoplasma hyopneumoniae infection. The B cell epitope of the present invention can be functionally expressed on the surface of an inert carrier S9 bacterium. Based on the LN7-antibody direct-mediated agglutination test detection method established by LN7 functionally expressed on the surface of the S9 bacterium, LN7 specifically recognizes only Mycoplasma hyopneumoniae infection serum antibodies and vaccine immune serum antibodies containing the GAPDH antigen, generating a specific binding reaction, while having no cross-reaction or non-specific binding reaction with other mycoplasma pathogens, porcine respiratory disease pathogen infection serum, and vaccine immune serum. This diagnostic method is expected to become an important technical means for the accurate diagnosis of Mycoplasma hyopneumoniae infection and the monitoring and evaluation of the immune effect of Mycoplasma hyopneumoniae vaccine.

This invention relates to the field of biomedical technology, providing a rabbit hemorrhagic disease virus (RHDV) type 2 fusion antigen protein and its applications. The fusion antigen protein comprises the RHDV2 VP60 protein sequence and an immunomodulatory CRM197 protein functional fragment, fused together via a linker peptide, and expressed and purified in an insect cell expression system. This invention improves the immunogenicity of the antigen protein by fusing the major capsid protein VP60 of RHDV2 with the immunomodulatory fragment of CRM197. Rabbit immunization experiments verified that the fusion antigen protein can induce high-titer specific antibodies, with serum antibody titers reaching 1:12800 and above, demonstrating good immunization effects. The fusion antigen protein of this invention can be used to prepare a vaccine for the prevention of RHDV2 infection, providing a new technical solution for the prevention and control of RHDV2 disease.

Disclosed herein are reagents for use in quality control assessment of antibody profiling platforms, such as biopanning and Digital Serology. Also disclosed herein are kits, method of manufacturing, and methods of using the same.

The application belongs to the field of animal bacteriology and molecular biology technology, and relates to preparation of a hybridoma cell and application thereof. A monoclonal antibody is prepared through intracellular Lawsonia intracellularis outer membrane protein OmpA expressed by prokaryotes, and IFA, WB and blocking ELISA and other immunological detection methods for detecting or diagnosing Lawsonia intracellularis are established through the expressed intracellular Lawsonia intracellularis outer membrane protein OmpA and the prepared monoclonal antibody. The blocking ELISA for detecting serum antibodies of Lawsonia intracellularis can detect serum samples of suspected Lawsonia intracellularis infection from different species, and does not exist cross reaction with positive serum of common pig pathogens, has high sensitivity and specificity, is good in repeatability, and is suitable for high-throughput detection of serum samples.

Lassa fever is a zoonotic viral hemorrhagic disease caused by Lassa virus (LASV), endemic in multiple west African countries where the multimammate rat is a main virus reservoir. The broad geographic range of the rodent host places a large population at risk of serious illness. LASV is also a bioterrorism threat. A LASV vaccine is an unmet need. Applicants advance a vaccine based on a live vesicular stomatitis virus vector for immunizing against the LASV surface glycoprotein. Preclinical evaluation exhibited high efficacy in cynomolgus macaques. The animals developed serum antibodies that could mediate antiviral effector functions including direct neutralization of virus infectivity. This vaccine candidate elicited robust protective immunity in cynomolgus macaques that prevented development of detectable viremia and disease for at least 1 year after vaccination. These protective immune response features provide valuable data for comparison to an ongoing phase 1 clinical trial and planned phase 2b / 3 clinical trials.

The invention discloses a B cell conservative epitope peptide LF7 of human papilloma virus (HPV) L1 protein and application of the B cell conservative epitope peptide LF7 in HPV infection early-stage antibody detection. An agglutination test antibody detection system based on direct mediation of an LF7-antibody is constructed by displaying and expressing LF7 on the surface of an inert vector bacterium S9H, the detection system can specifically detect and monitor HPV infection early infectious antibodies and does not have cross reaction with other pathogen infection positive serum antibodies, and by taking the inert vector bacterium S9H which does not express LF7 epitope peptide as a contrast, the detection system can detect and monitor HPV infection early infectious antibodies and does not have cross reaction with other pathogen infection positive serum antibodies. The reaction with infectious antibodies is also avoided. In the detection of a positive serum sample at the early stage of HPV infection, the detection positive rate is 100%. The antibody detection system has the advantages of strong specificity and high sensitivity, can quickly and accurately detect the HPV infection early infectious antibody, and can quantitatively determine the titer of the antibody. The invention provides efficient and reliable technical support for early accurate diagnosis of HPV infection, and has important clinical application value.

The present invention discloses an EBV whole protein chip and a preparation method and application thereof, wherein the EBV whole protein chip comprises a substrate and 85 EBV proteins connected thereto. The EBV whole protein chip prepared by the present invention can be used to screen EBV proteins that interact with serum antibodies, and has the characteristics of global and high-throughput screening, simplifies the research design and operation process, and significantly improves the research efficiency.

The present application relates to the technical field of novel coronavirus, and particularly relates to a novel coronavirus trivalent DNA vaccine and a preparation method. The trivalent DNA vaccine takes a recombinant plasmid carrying a nucleotide sequence as shown in SEQ ID NO: 1 or 2 as an active ingredient. The trivalent DNA vaccine provided in the present application can express a fusion antigen protein of three kinds of antigen genes of the novel coronavirus in eukaryotic cells, and produce serum IgG antibodies against the three kinds of antigens of the novel coronavirus. In addition, the novel coronavirus trivalent DNA vaccine provided in the present application can immunize mice to produce T cells secreting various cytokines, and has a stronger T cell immune protection effect.

The invention discloses a mycoplasma hyopneumoniae ELISA (enzyme-linked immunosorbent assay) serum antibody detection method. The mycoplasma hyopneumoniae ELISA serum antibody detection method comprises the following steps: coating an ELISA plate with an antigen, sealing, incubating serum, incubating a second antibody and developing a substrate, wherein the antigen is purified Mhp336 protein. The invention also discloses a system for diagnosing swine mycoplasmal pneumonia. The mycoplasma hyopneumoniae ELISA serum antibody detection method disclosed by the invention has the characteristics of good repeatability, high specificity and high sensitivity.

The invention relates to the technical field of poultry immunology, in particular to a universal riemerella anatipestifer monoclonal antibody, a detection kit and application of the universal riemerella anatipestifer monoclonal antibody. Specifically, the invention successfully designs the universal riemerella anatipestifer monoclonal antibody, and the detection kit developed based on the monoclonal antibody can rapidly detect the level of the riemerella anatipestifer serum antibody in a sample, and has high sensitivity and strong specificity. The riemerella anatipestifer strain has no cross reactivity with avian enterococcus faecalis, avian salmonella typhimurium, avian escherichia coli and avian salmonella enteritidis, so that the riemerella anatipestifer strain can be applied to rapid screening, epidemiological monitoring and vaccine immune effect evaluation of riemerella anatipestifer infection, and has a wide application prospect.

The present invention provides a method for preparing a serum antibody affinity separation material, which comprises washing an agarose matrix, epoxy activation reaction, and coupling with a recombinant Streptococcus Protein G (SPG) with agarose as a matrix to prepare the serum antibody affinity separation material; or washing an agarose matrix, epoxy activation reaction, amination reaction of the activated agarose, and coupling to prepare the serum antibody affinity separation material. Also provided is an application of the serum antibody affinity separation material prepared by the present invention, which is used for adsorbing immunoglobulin G (IgG). The serum antibody affinity separation material prepared by the present invention does not need to use the classic but highly toxic cyanogen bromide as a coupling reagent, and has good biosafety.

The invention discloses application of plant lactobacillus DPUL-F232 and capsular polysaccharide thereof in food allergy, and belongs to the technical field of microorganisms. The lactobacillus plantarum DPUL-F232 capsular polysaccharide disclosed by the invention can be used for relieving allergic symptoms and weight change of rats; the spleen index and rectum temperature change are reduced; the levels of serum antibodies IgE, IgG1 and IgG2a are reduced; mast cell protease and histamine levels are reduced; regulating the levels of cell factors IFN-gamma, IL-4, IL-10, IL-6 and IL-17A of serum and splenocytes; the heat-inactivated lactobacillus plantarum DPUL-F232 capsular polysaccharide is very high in stability, can be stored for a long time, is suitable for preparing anti-allergic foods or medicines, and has a relatively good application prospect.

The invention provides a lysosome targeting organic polymer nanoparticle vaccine as well as a preparation method and application thereof. The organic polymer nanoparticle vaccine can target lysosome, promote maturation of antigen presenting cells and secretion of proinflammatory cytokines, improve serum antibody titer and enhance immune protection effect on pathogen infection. The invention also provides an organic high-molecular polymer for preparing the organic high-molecular nanoparticle vaccine, and the organic high-molecular polymer can accurately co-deliver adjuvants and antigens, has good biocompatibility and degradability, and can reduce immunogenicity and side effects.

The invention discloses an immune effect evaluation method of a cysticercus cellulosae recombinant TPx protein vaccine, and belongs to the technical field of biological medicines. The vaccine takes recombinant thioredoxin peroxidase (TPx) from cysticercus cellulosae as an immunogen, and an adjuvant (such as a Freund's adjuvant) can be optionally supplemented to enhance the immune effect. A mouse is immunized through subcutaneous injection of the vaccine, it is proved that the vaccine can effectively induce an organism to generate humoral immune response and cellular immune response, specifically, the levels of serum IgG, IgG2a and IgG1 antibodies are remarkably increased, the proliferation capacity of splenic lymphocytes is enhanced, and the secretion levels of Th1 type cell factors (TNF-alpha and IL-12) and Th2 type cell factors (IL-13 and IL-10) are remarkably increased. The invention provides a new candidate vaccine for prevention of cysticercosis, and has important application value.

The invention discloses a protein composition for serum antibody detection and application of the protein composition in early diagnosis of lung cancer, and belongs to the technical field of biological medicine. The protein composition comprises an antigen protein TGIF2LY and any one or more of antigen proteins CHIC2, RAC3, HRAS, CDK9, HK2, TXNDC2, TEX264 and N4BP1, on the basis of improving the sensitivity, the same average value of sensitivity and specificity can be basically achieved, particularly, the protein composition comprises the antigen proteins TGIF2LY and CHIC2, the sensitivity is 64%, the specificity is 95%, and the average value of sensitivity and specificity is 79%. Besides, the antigen proteins CHIC2, RAC3, CDK9 and HK2 are newly screened antigen proteins and can be used for detecting the IgG antibody of the lung cancer in serum so as to be used for early diagnosis of the lung cancer, and the sensitivity, the specificity and the average value of the sensitivity and the specificity are basically the same as those of a single antigen protein in the prior art and even better than those of the single antigen protein in the prior art.

The invention discloses a novel duck reovirus RD240407A with strong pathogenicity as well as preparation and application of an egg yolk antibody of the novel duck reovirus RD240407A. Specifically, the preservation number of the novel duck reovirus RD240407A is CGMCC (China General Microbiological Culture Collection Center) No. 46196. The invention also provides an inactivated vaccine and an egg yolk antibody prepared from the strain. The egg yolk antibody prepared from the novel duck reovirus RD240407A has a good immune effect on disease treatment and prevention before disease of ducklings and goslings. Compared with a serum antibody in the prior art, the preparation method of the egg yolk antibody is simple, blood sampling is not needed, and the egg yolk antibody can be obtained by only collecting and purifying immune hyperimmune egg yolk; the preparation method is simple and convenient, and is suitable for large-scale production, popularization and application.

The present invention discloses a fusion tandem polypeptide antigen for detecting antibodies against African horse sickness virus. The present invention provides a polypeptide or a set of polypeptides. The polypeptide is eVP7-1; the set of polypeptides consists of eVP2-1, eVP5-1 and eVP7-1 or consists of eVP5-1 and eVP7-1; the eVP2-1 is the polypeptide of SEQ ID No.1; the eVP5-1 is the polypeptide of SEQ ID No.4; the eVP7-1 is the polypeptide of SEQ ID No.7. The antibody detection kit prepared with BSA-eVP2-1 + BSA-eVP5-1 + BSA-eVP7-1 or BSA-eVP5-1 + BSA-eVP7-1 as the coating antigen has strong specificity, high sensitivity, high accuracy, simple and rapid operation, and is suitable for rapid and large-scale screening and detection of serum antibodies against African horse sickness virus by grass-roots veterinary departments at all levels and entry-exit inspection and quarantine bureaus.

The invention belongs to the technical field of high-throughput detection, and discloses a high-throughput sequencing library as well as a construction method and application thereof. The invention provides a construction method of a phage display library and an obtained T7 phage display library. According to the phage display library, a metagenome sequencing technology and a phage display technology are combined, and the phage display library has the advantages of high throughput, low cost, ultra-large capacity and the like. The invention further provides a construction method of the high-throughput sequencing library and the obtained high-throughput sequencing library. According to the present invention, the phage display library is constructed, the immunoprecipitation and the serum are adopted to co-incubate and capture the antibody, the high-throughput sequencing library is constructed, the sequencing is performed, and the serum antibody information is obtained through the bioinformatics analysis so as to achieve the high-throughput serological test, the specificity is strong, and the sensitivity is high.

The application belongs to the technical field of biology and specifically relates to a SAT1 type foot-and-mouth disease virus monoclonal antibody 5G10, a kit and a detection method. The single-chain antibody 5G10 comprises a heavy chain variable region and a light chain variable region, the amino acid sequence of the heavy chain variable region is shown in SEQ ID No. 6, and the amino acid sequence of the light chain variable region is shown in SEQ ID No. 7. The titer is greater than 1:128000, the heavy chain is IgG1 type, and the light chain is lambda type. The antibody can specifically combine with VP1 recombinant protein of the SAT1 type foot-and-mouth disease virus and has high specificity. The application of the monoclonal antibody 5G10 in a foot-and-mouth disease virus SAT1 type blocking ELISA antibody detection method is provided, and convenient and effective technical reserves are provided for border port detection and animal serum antibody screening of the SAT type foot-and-mouth disease.