63 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 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 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 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 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 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 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.

The invention provides an orf virus double-antigen self-assembled nanoparticle, the nanoparticle is a fusion protein containing orf virus protein and ferritin, and the orf virus protein comprises orf virus protein ORFV 050 and orf virus protein ORFV 086. The ORFV050-ORFV086-Fel fusion protein prepared by the invention is self-assembled into nanoparticles in vitro, and can induce the generation of high-titer serum antibodies and cell factors such as IFN-gamma, TNF-alpha and IL-6 of an organism, so that the ORFV050-ORFV086-Fel fusion protein can be used for preparing vaccines for preventing and / or treating orf. Based on ORFV 050-ORFV086-Fir fusion protein, the self-assembled protein nanoparticle vaccine provided by the invention is safe and effective, can induce immune mice to generate lasting immune protection, has good immune efficacy, and can be clinically used for prevention and / or treatment of orf virus infection.

The kit for systemic lupus erythematosus detection comprises a lower box body, an upper box body, an upper placement groove, a lower placement groove, a strip-shaped air bag, a limiting rod, an inflation bag, an exhaust bag and a first flow dividing pipe, the upper box body is arranged above the lower box body in an overturning mode, the upper placement groove is formed in the inner side of the upper box body, the lower placement groove is formed in the inner side of the lower box body, and the strip-shaped air bag is arranged in the lower placement groove. The lower end face of the lower box body is provided with a limiting rod, an inflation bag and an exhaust bag, the lower end face of the lower box body is provided with an air exhaust bag, and a first flow dividing pipe is embedded in the lower end face of the lower box body, so that the problem that the protection effect of an original kit on a serum antibody reagent is poor is solved; the serum antibody reagent placing box is reasonable in structure, after a serum antibody reagent is placed, the strip-shaped air bag in the lower placing groove can be inflated by pulling the inflation bag so as to protect the serum antibody reagent, and the serum antibody reagent placing box is simple and convenient to use and high in practicability.

The present invention relates to a method for enhancing gene therapy by using effective degradation or digestion of serum antibodies in a subject and / or inhibition or reduction of effector functions of serum antibodies in a subject. More specifically, the subject does not have a detectable antibody that specifically binds to gene therapy, or has a low titer neutralizing antibody (NAb) that specifically binds to gene therapy or a low titer total antibody (TAb) that specifically binds to gene therapy. The methods can increase the serum half-life of the therapy and indicate that lower gene therapy doses can be administered than other methods.

The present invention discloses an indirect ELISA method for detecting the level of ApxI toxin antibody in porcine serum samples, and the indirect ELISA method comprises the following steps: Step S1: Prepare the antigen; Step S2: Determine the conditions of the ELISA method; Step S3: Measure the positive standard product to obtain the average value X and the standard deviation S of OD 450nm , and determine the positive and negative critical values of the indirect ELISA method; Step S4: Measure the sample to be tested. By optimizing conditions such as the antigen coating concentration, serum dilution ratio and action time, enzyme-labeled secondary antibody dilution concentration and action time, antigen-antibody binding time, blocking time, substrate chromogenic time, etc., the present invention can effectively detect the serum antibody after immunizing with the APP vaccine.

The invention provides a quantitative detection method of a porcine reproductive and respiratory syndrome serum antibody, and belongs to the technical field of veterinary immunological detection.The method comprises the steps that S1, a serum sample is collected and pretreated; s2, antibody marker extraction and characteristic parameter extraction; s3, feature processing and antibody signal enhancement based on four-algorithm fusion; and S4, quantitative calibration and antibody concentration output. Through closed-loop design of multi-algorithm fusion, accurate quantitative detection of the porcine reproductive and respiratory syndrome serum antibody is realized.

The present invention relates to the technical field of in vitro detection reagents, and specifically to a porcine circovirus antibody detection kit based on an indirect immunofluorescence method and its application. The porcine circovirus Cap antigen provided by the present invention can simultaneously bind to serum antibodies induced by two genotypes, TTSuV1 and TTSuV2, and has high specificity. Based on the Cap antigen and the IFA method, the present invention establishes a serological detection kit and detection method for TTSuV antibodies, which can simultaneously detect serum antibodies induced by two genotype viruses, TTSuV1 and TTSuV2, and has good specificity, stability and repeatability, effectively avoiding the problems encountered in the establishment process of the ELISA method, and can better meet the needs of TTSuV serological clinical detection, and has good application prospects.

This invention relates to the field of avian immunology, and more particularly to a universal monoclonal antibody against Riemerella anatipestifer, a detection kit, and their applications. Specifically, this invention successfully designed a universal monoclonal antibody against Riemerella anatipestifer, and the detection kit developed based on this monoclonal antibody can rapidly detect the level of Riemerella anatipestifer serum antibodies in samples with high sensitivity and strong specificity. Furthermore, it shows no cross-reactivity with Enterococcus faecalis, Salmonella typhi, Escherichia coli, and Salmonella enteritidis, thus it can be used for rapid screening, epidemiological monitoring, and evaluation of vaccine immunization efficacy against Riemerella anatipestifer infection, with broad application prospects.

The invention discloses a B cell conservative epitope peptide of enterotoxigenic Escherichia coli V-type secretion system effect protein EatA and application thereof, and particularly relates to application in ETEC specific antibody diagnosis. The B cell conservative epitope peptide disclosed by the invention can be functionally displayed and expressed on the surface of an inert carrier S9H thallus, PT8 is displayed and expressed on the surface of the S9H thallus, and an agglutination test directly mediated by the PT8 and specifically recognized by an antibody is established, so that only ETEC pathogen infectious antibodies are specifically recognized, other pathogenic escherichia coli can be identified and diagnosed, and the B cell conservative epitope peptide has a good application prospect. And no cross reaction with serum antibodies of other infectious diseases exists. According to the method, the specific serum antibody can be detected 7 days after a Balb / c mouse is infected by the ETEC strain H10407. The diagnosis method is high in specificity, good in sensitivity, convenient to operate and intuitive in result judgment, and is expected to become a technical means for visual on-site accurate and convenient diagnosis of ETEC infection.

The invention discloses a preparation method and application of a colloidal gold test strip for detecting a group A rotavirus antibody. The gold-labeled protein used by the test strip is prokaryotically expressed and soluble RVA-VP6-3P recombinant protein, and the protein has rich group-specific epitopes, so that the specificity and sensitivity of the gold-labeled protein are ensured, the economic cost for obtaining and purifying the target protein is reduced, and a foundation is laid for establishment, popularization and application of an antibody detection method. The RVA-VP6-3P is labeled with colloidal gold, the detection line is coated with SPA, the quality control line is coated with a monoclonal antibody 1M1F9 capable of specifically recognizing the RVA-VP6-3P, and then the test strip is assembled. The test strip has no cross reactivity with positive serum antibodies of other common swine disease viruses, has the characteristics of convenience in operation, less time consumption, high sensitivity, strong specificity and the like, can complete the detection of the group A rotavirus antibody in 5-15 minutes, and is convenient for immediate use in a clinical first line.