92 results about "Titin Antibody" patented technology

The invention belongs to the technical field of biochemical engineering, and particularly relates to an amitriptyline hapten and a preparation method thereof, an amitriptyline antigen and a preparation method thereof, an antibody, a kit and application. The amitriptyline hapten has a structural formula shown as a formula (I) or a formula (II). Compared with an existing structure, the amitriptyline hapten prepared by the invention has a better immune effect. The obtained antibody is high in titer, strong in specificity and high in affinity, the LOD of amitriptyline by using an Elisa method established by the antibody is 0.013 ng / mL, the IC50 of amitriptyline is 0.865 ng / mL, the quantitative detection range is 0.07-470.3 ng / mL, the detection sensitivity is high, and the linear range is wide.

The invention provides an antigen epitope peptide related to connexin and application of the antigen epitope peptide. The connexin antigen epitope peptide disclosed by the invention is selected from (1) a polypeptide with an amino acid sequence as shown in SEQ ID NO: 14; and (2) a polypeptide which is derived from (1) by substituting, deleting or adding 1-2 amino acids in the amino acid sequence of SEQ ID NO: 14 and retains the binding capacity with a connexin antibody. According to the present invention, the new connexin antigenic peptide fragment sequence is identified for the first time, such that the existing autoantigen epitope map is expanded, and more importantly, the molecular basis is provided for the development of the clinical detection method with high diagnosis sensitivity and high specificity, such that the pathological mechanism of MG can be further improved, and the application prospect is broad. And new auxiliary diagnosis means and treatment targets are provided for antibody negative MG patients.

The invention discloses an anti-CD3D antibody and application thereof.The antibody comprises a heavy chain variable region and a light chain variable region, the amino acid sequence of CDR-H1 of the heavy chain variable region is shown as SEQ ID NO: 4, the amino acid sequence of CDR-H2 of the heavy chain variable region is shown as SEQ ID NO: 5, and the amino acid sequence of CDR-H3 of the heavy chain variable region is GDL; the amino acid sequence of the CDR-L1 of the light chain variable region is as shown in SEQ ID NO: 6, the amino acid sequence of the CDR-L2 of the light chain variable region is as shown in SEQ ID NO: 7, and the amino acid sequence of the CDR-L3 of the light chain variable region is as shown in SEQ ID NO: 8. The anti-CD3D antibody is a rabbit monoclonal antibody, the obtained antibody is high in titer, clear in specific positioning in immunohistochemical staining and free of non-specific background staining, the antibody is wide in application, and the antibody can be used in pathological immunohistochemical staining experiments and can also be used in Western blot and flow cytometry experiments.

The invention discloses an influenza C virus HEF protein polyclonal antibody preparation method, which comprises: carrying out signal peptide prediction, antigen epitope screening and hydrophobicity analysis through bioinformatics, constructing a recombinant prokaryotic expression vector pET32a-HEF by using a seamless cloning technology, and efficiently expressing the HEF protein in Escherichia coli to obtain the influenza C virus HEF protein polyclonal antibody. The inclusion body protein yield is improved by optimizing conditions; and purifying by nickel column affinity chromatography to obtain high-purity recombinant protein, emulsifying the high-purity recombinant protein and a Freund's adjuvant, immunizing a Japanese white rabbit, immunizing for three times, and collecting high-titer serum. The method breaks through detection limitation caused by antigen tag concealment in a eukaryotic expression system, the obtained polyclonal antibody is high in specificity, the eukaryotic expression HEF protein can be accurately recognized, and a key tool is provided for ICV virus-like particle identification, vaccine research and development and diagnosis technologies. Compared with a traditional method, the scheme is simple and convenient to operate, low in cost and high in antibody titer, and has remarkable application value.

The invention discloses an application of an AP2-F2 protein polyclonal antibody as a cryptosporidium female gamete positioning tag antibody. The AP2-F2 protein of cryptosporidium parvum is subjected to truncated prokaryotic expression, protein is purified to immunize an ICR mouse, a polyclonal antibody is prepared, ELISA and WB detection of antibody titer are carried out to prove that the polyclonal antibody is effective, then indirect immunofluorescence detection of the antibody is carried out to determine that the polyclonal antibody is located on the female gametes of cryptosporidium parvum, and the polyclonal antibody can be used as a label to locate the female gametes of cryptosporidium parvum. The method plays an important role in localization research of unknown protein of cryptosporidium.

The preparation and evaluation method comprises the following steps: 1, extracting a heavy-chain DNA fragment and a light-chain DNA fragment, carrying out recombinant transformation by using an overlapping PCR method to construct an scFv gene, 2, constructing an expression vector, culturing the expression vector, carrying out sequencing identification to obtain an scFv bacterial solution, recombining the scFv bacterial solution with a mouse Fc fragment, and identifying a sequence to obtain an MN8 bacterial solution, the method comprises the following steps: 1, preparing an MN8 bacterial liquid, 2, extracting the MN8 bacterial liquid, 3, carrying out inoculated culture on the MN8 bacterial liquid to obtain an MN8 genetically engineered antibody bacterial liquid, 4, extracting MN8 genetically engineered antibody bacterial liquid plasmids and carrying out transfection expression purification, and 5, carrying out antibody titer and thermal stability determination on the MN8 genetically engineered antibody. The scFv single-chain antibody obtained by recombination according to the method has the advantages of small molecular weight, high penetrability and the like, the stability and the sensitivity of an immunoassay method of the scolosporins can be improved, and a certain basis is provided for rapid determination of the scolosporins.

This invention discloses a hybridoma cell line that secretes anti-polystyrene monoclonal antibodies and its applications. The hybridoma cell line includes PS-8, classified as Hybridoma cell line PS-8. The accession number for PS-8 is CCTCC NO: C2024144, the accession date is May 10, 2024, and the depositary institution is the China Center for Type Culture Collection (CCTCC), located at Wuhan University, No. 299 Bayi Road, Wuchang District, Wuhan City, Hubei Province. The hybridoma cell line provided by this invention can stably secrete anti-polystyrene monoclonal antibodies with high titer, good specificity, and high sensitivity, making it suitable for detection in reagent kits and possessing potential application value.

The invention discloses an African swine fever virus vaccine as well as related products and application thereof, and relates to the field of vaccines. The saRNA vaccine of the African swine fever virus provided by the invention adopts an optimized antigen sequence and a saRNA skeleton, and compared with a traditional African swine fever virus vaccine, the saRNA vaccine of the African swine fever virus has the advantages of being early in antibody production period, good in safety, simple in immune procedure, higher in antibody titer and the like.

The invention belongs to the technical field of vaccine preparation, and particularly relates to a method for suspension culture of rabies virus. MDCK-siat7e-C is used as a cell matrix for suspension culture of the rabies virus, experiments prove that the rabies virus can be subcultured in the cell, the virus titer can reach 7-81gFFU / ml, and through mouse immunogenicity inspection, the protection index is greater than 100, and the antibody titer reaches 67IU / ml after mice are immunized. The method for suspension culture of the rabies virus by the cell strain provides reference for suspension culture of the rabies virus.

The application provides a preparation method of polysaccharide vaccine diluent and application to chicken newcastle disease vaccine, and belongs to the technical field of biopharmacy.The application provides a south five canes polysaccharide vaccine diluent and a preparation method thereof, and solves the problems of low uniformity of antibody titer, stress reaction and allergic reaction of the body after inoculation, and influence on the immunization effect of the vaccine caused by the physiological saline or the phosphate buffer solution diluent for diluting the vaccine.The south five canes polysaccharide vaccine diluent can be applied to animal vaccines, including chicken newcastle disease vaccine, and solves the problem that there is no polysaccharide diluent for the chicken newcastle disease vaccine in the prior art.

The invention relates to the field of biological products, and particularly discloses a goose astrovirus, a goose astrovirus egg yolk antibody as well as a preparation method and application of the goose astrovirus egg yolk antibody. The invention relates to a goose astrovirus, which is named as RD240629A, is preserved in the China General Microbiological Culture Collection Center (CGMCC), the preservation number is CGMCC No: 46197, and the preservation date is September 23, 2024. The goose astrovirus is named as RD240629A, and the goose astrovirus is named as RD240629A and is preserved in the China General Microbiological Culture Collection Center (CGMCC). The goose astrovirus can be used for preparing the egg yolk antibody for preventing gout of goslings, the egg yolk antibody is good in safety, no local or whole-body adverse reaction caused by antibody injection occurs, infection of the novel goose astrovirus can be effectively prevented and / or treated, and the anti-gout effect is good. The prepared egg yolk antibody is evaluated by adopting antibody titer determination and a challenge protection test, the titer of the egg yolk antibody is not lower than 1: 256, and the egg yolk antibody has a good commercial development prospect.

The invention discloses a polyclonal antibody of panda Rbp7 protein as well as a preparation method and application thereof, and belongs to the technical field of antibodies. The first to nineteenth amino acid sequences of the panda Rbp7 protein are used as antigen polypeptides, and are specifically shown as SEQ ID No.1. After chemical synthesis or exogenous expression purification, a Balb / C mouse is immunized, and the polyclonal antibody is obtained through repeated immunization, serum titer detection and affinity chromatography purification. The titer of the antibody is not lower than 1 / 128000, the panda Rbp7 recombinant protein can be efficiently and specifically detected through WB and ELISA, the blank of a protein detection tool is filled, and reliable technical support is provided for research on the functional mechanism of the panda Rbp7 protein.

The invention discloses design and synthesis of a hapten of cyromazine, development of a cyromazine artificial antigen and a cyromazine antibody, and application of the cyromazine artificial antigen and the cyromazine antibody in the field of food safety rapid detection. The aminocarboxylic acid modified cyromazine hapten product is prepared through the steps of cyromazine hapten design, connecting arm introduction, synthesis, purification and the like. The cyromazine hapten with the structure is coupled with keyhole limpet hemocyanin to form an artificial immune antigen with immunogenicity, and is coupled with chicken ovalbumin to form an artificial competitive antigen. Tests show that the artificial immune antigen has high immunocompetence and can stimulate an animal immune system to generate an anti-cyromazine antibody, and the titer of an immune Blab / c mouse serum antibody is 16000. The IC50 of an enzyme-linked immunosorbent assay (ELISA) detection method established by the cyromazine artificial competitive antigen and the anti-cyromazine antibody is 5.3 [mu] g / L, and the detection sensitivity of an established gold-labeled immunochromatography detection method reaches 50 [mu] g / L.

The invention provides a freeze-dried universal platelet rich plasma (PRP) preparation and a preparation process thereof, aiming at solving the problems that the existing PRP is limited in blood type matching, difficult in biological activity preservation, short in validity period and the like. According to the preparation, 18-19% of the type A, 7-8% of the type B and 73-74% of the type AB are mixed with PRP of different blood types and freeze-dried. Tests show that agglutination does not exist in cross blood matching, the titers of the anti-A antibody and the anti-B antibody are both lower than 1: 2, and blood group reaction is effectively avoided. Freeze-drying causes little damage to growth factors, still has biological activity within 24 months, and has a good cell proliferation test effect. Animal experiments show that wound healing can be remarkably promoted. In addition, the culture medium is good in stability in the environment of 4 DEG C, and the moisture, growth factors and the sterile state all meet the requirements. The preparation disclosed by the invention has the advantages of obvious universal advantages, good biological activity preservation and strong stability, and provides a better choice for clinical treatment.

The invention discloses a halogen modified hapten for enhancing antibody titer and affinity as well as a preparation method and application of the halogen modified hapten, and belongs to the technical field of biochemical engineering. According to the method, an amantadine structure is innovatively and chemically modified, various halogens such as fluorine, chlorine and bromine are introduced, the AMA halogen modified hapten shown in the formula (I) is formed, and the traditional'most similar 'design principle is broken through. Compared with an unmodified hapten, the hapten modified by halogen in the invention has the advantages that the titer of the polyclonal antibody on amantadine can be obviously improved by 3.4-14 times; wherein the haptens A5 and A6 can obviously improve the titer of the monoclonal antibody to amantadine by 3-18 times, and the affinity is improved by 10.8-16.1 times. The invention provides a new thought and a new method for the rational design of the hapten of the small molecule compound and the preparation of the high-affinity antibody, and the related halogen modification type and quantity strategy can also provide valuable reference for the design of the hapten of other small molecule compounds.

The invention discloses a neutralizing antibody with an AQP3 channel blocking function as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The neutralizing antibody can be specifically combined with an extracellular domain of AQP3, after combination, a water channel and a small molecule transport function of the AQP3 are blocked through steric hindrance and conformation change, and meanwhile, antigen combination specificity is reserved. The preparation method is realized through'recombinant antigen-immunization-B cell screening culture-recombinant antibody expression and affinity purification-ELISA detection ', and the functions of the antibody are ensured. The antibody can be efficiently combined with AQP3 positive cells, is strong in targeting property and high in specificity (the serum titer reaches 1: 1024 * 10, and the purified antibody titer is greater than or equal to 1: 128 * 10), and can be independently used or combined with a nano-carrier for treating related diseases, so that the technical bottleneck that a common AQP3 antibody can only be combined and cannot efficiently play a blocking role is solved.

The present invention relates to the field of immunoassay technology and provides a bifenthrin hapten, antigen, antibody, preparation method, and application thereof. The bifenthrin hapten provided by the present invention completely retains the molecular structure of bifenthrin. After coupling with a carrier protein, the immunogenicity of the resulting antigen is significantly improved, and the prepared antibody has stronger specificity and higher sensitivity. The results of the examples show that the bifenthrin hapten provided by the present invention is coupled with a carrier protein, and the resulting antigen, after injection into mice for immunization, produces an antibody titer of 1×10 4 , half inhibitory concentration (IC 50 ) is about 20ppb, and the minimum detection limit of colloidal gold for bifenthrin in dry tobacco leaf samples is about 1.25μg / g.

The present invention discloses a rabies inactivated vaccine diluent and its preparation method and application, belonging to the technical field of vaccine diluents. The invention provides an inactivated vaccine diluent to solve the technical problems of low antibody titer, long production time and short antibody maintenance time after animal inactivated vaccine immunization. The invention provides a rabies inactivated vaccine diluent, which is composed of the following components: phosphate buffer, MONTANIDE TM GEL 02 PR adjuvant, canine interferon, and ricin B chain protein. This diluent can significantly increase the rabies neutralizing antibody titer after rabies inactivated vaccine administration.

The invention discloses a hybridoma cell strain secreting an anti-PS monoclonal antibody and application of the hybridoma cell strain, the hybridoma cell strain comprises a hybridoma cell strain PS-17, the hybridoma cell strain PS-17 is preserved in the China Center for Type Culture Collection, and the preservation number of the hybridoma cell strain PS-17 is CCTCC NO: C2024145. The passage cell strain of the hybridoma cell provided by the invention can stably secrete the anti-polystyrene protein monoclonal antibody, the secreted antibody is high in titer, good in specificity and high in sensitivity, and the passage cell strain can be applied to kit detection and has potential application value.

The invention relates to the technical field of antibody titer digitization analysis, in particular to a brucellosis antibody titer digitization analysis system based on a spectrophotometric method.According to the brucellosis antibody titer digitization analysis system, a full-time reaction kinetics absorbance curve is obtained through a data acquisition module; a signal decomposition module is utilized to decouple the signal into a low-frequency trend component and a high-frequency detail component; further identifying a reaction kinetics phase state through a low-frequency trend analysis module to lock an effective reaction interval and generate first-dimension analysis data, and controlling a high-frequency detail analysis module to generate second-dimension analysis data by taking the first-dimension analysis data as a time domain gating basis; finally, the comprehensive evaluation module constructs a coupling weight model based on the reaction kinetics phase state, and fuses the two-dimensional data to generate a comprehensive antibody activity evaluation value; according to the process, a self-adaptive analysis mechanism conforming to a reaction evolution rule is constructed, and accurate determination of the titer of the antibody is realized.

The preparation and evaluation method comprises the following steps: 1, extracting a heavy-chain DNA fragment and a light-chain DNA fragment, carrying out recombinant transformation by using an overlapping PCR method to construct an scFv gene, 2, constructing an expression vector, culturing the expression vector, carrying out sequencing identification to obtain an scFv bacterial solution, recombining the scFv bacterial solution with a mouse Fc fragment, and identifying a sequence to obtain an MN8 bacterial solution, the method comprises the following steps: 1, preparing an MN8 bacterial liquid, 2, extracting the MN8 bacterial liquid, 3, carrying out inoculated culture on the MN8 bacterial liquid to obtain an MN8 genetically engineered antibody bacterial liquid, 4, extracting MN8 genetically engineered antibody bacterial liquid plasmids and carrying out transfection expression purification, and 5, carrying out antibody titer and thermal stability determination on the MN8 genetically engineered antibody. The scFv single-chain antibody obtained by recombination according to the method has the advantages of small molecular weight, high penetrability and the like, the stability and the sensitivity of an immunoassay method of the scolosporins can be improved, and a certain basis is provided for rapid determination of the scolosporins.

The invention is applicable to the technical field of gene engineering, and is characterized in that a streptococcus zooepidemicus ATCC 35246 strain is used as a parent strain, and a pSET4S temperature-sensitive suicide gene knockout plasmid is utilized to knock out pyrroline-5-carboxylic acid reductase gene (proC), so that a streptococcus zooepidemicus gene deleted strain ATCC 35246 delta proC is obtained. The deletion strain has low toxicity, and when the mutant strain is used for immunizing a mouse in a large dose of viable bacteria, the titer of an antibody in the mouse can be obviously increased, and targeted immune protection is successfully established; the ATCC 35246 delta proC is convenient to prepare, low in cost and simple to culture, does not need other complex treatment when being used as a vaccine, and can be used after being adjusted to a corresponding concentration after bacteria washing. A result of an immune experiment on a mouse shows that the vaccine can effectively stimulate the immune level of an animal body and resist streptococcus zooepidemicus infection, and the deletion strain is high in safety and remarkable in immune protection effect in the mouse and has the potential of serving as a streptococcus zooepidemicus live vaccine.

This invention discloses monoclonal antibodies that recognize linear B-cell epitopes in the conserved extracellular domain of pseudorabies virus (PRV) gE protein and their applications. Using recombinant PRV gE protein extracellular domain protein as an immunogen, hybridoma cell lines were prepared. Combining dot blot and peptide scanning techniques, two monoclonal antibodies, 4E5D7 and 8F6B2, were screened to recognize the 69-93aa region of the conserved extracellular domain of the PRV gE protein. Both antibodies achieved a titer of 1:2048000 and specifically recognized the native PRV wild-type gE protein, showing no cross-reactivity with gE gene-deleted vaccines. The monoclonal antibodies of this invention fill the gap in detection reagents targeting the precise epitopes of the 69-81aa and 79-93aa extracellular domains of the PRV gE protein, thus improving the linear B-cell epitope map of the PRV gE protein. These monoclonal antibodies can be used for research on the pathogenesis of PRV and the development of specific detection reagents for PRV wild-type strains, possessing significant application value and translational prospects in basic scientific research, veterinary clinical testing, and animal disease prevention and control.

The present invention relates to the technical field of immunoassay, and provides a pendimethalin hapten, antigen, antibody, preparation method and application thereof. The present invention introduces an active group to the 2-methyl group of pendimethalin. After the obtained pendimethalin hapten is conjugated with a carrier protein, it has high immunogenicity. After immunizing a host animal, antibodies with high titer and high sensitivity can be produced. The pendimethalin hapten of the present invention is obtained by three steps of nitration reaction, substitution reaction and condensation reaction using 2-methyl-4-chlorobenzylamine as a raw material. The preparation method provided by the present invention is simple to operate, has mild conditions, easily available raw materials, and the product has high yield and high purity. The results of the examples show that after the pendimethalin antigen of the present invention is injected and immunized in mice, the antibody titer is 1.0×10 4 , and the half inhibitory concentration (IC 50 ) is about 0.5 ppb. The minimum detection limit of the colloidal gold reagent for pendimethalin in dry tobacco leaf samples is 10 ng / g.

The application provides an anti-glycocholic acid antibody and application thereof, and specifically, the application immunizes BALB / C mice with a glycocholic acid antigen, and more than ten positive monoclonal hybridoma cell strains are screened and obtained. Through performance determination, finally, 3 cell strains capable of specifically combining with glycocholic acid are obtained. Among them, the H15 antibody has high titer, good stability, and stable detection linearity, and can be used for clinical detection of glycocholic acid.

The present application relates to the field of immunoassay technology, and provides a myclobutanil hapten, antigen, antibody and preparation method and application. The present application introduces an active group at the meta position of the chlorine atom on the benzene ring of myclobutanil. Compared with the existing hapten, the hapten provided by the present application completely simulates the structure of myclobutanil, and enhances the immunogenicity of the antigen. The antigen provided by the present application has strong immunogenicity after being coupled with a carrier protein. After injection immunization of mice, the titer of the produced antibody is 5.0x10 4 , the half-inhibitory concentration (IC 50 ) is about 50 ppb, and the minimum detection limit of colloidal gold for myclobutanil in dry tobacco leaf samples is about 2.5 μg / g.

The present application relates to the technical field of bio-chemical engineering, and particularly relates to a halogen-modified hapten for enhancing antibody affinity, and a preparation method and application thereof. The present application provides a halogen-modified sulfonamide hapten for enhancing antibody affinity, which is obtained by adopting halogen-modified sulfonamides, and the antibody prepared by the halogen-modified sulfonamide hapten has relatively high titer and affinity. The present application provides a new and effective approach for preparing high-affinity sulfonamide antibodies.

The invention relates to the technical field of immunoassay, and provides a vomitoxin hapten, an antigen, an antibody, a preparation method and a light-activated chemiluminescence quantitative detection kit. The hapten is obtained by adopting cyclic dicarboxylic anhydride as a reactant and introducing an active group on a C atom of conjugated alkene of vomitoxin molecules through Friedel-Crafts acylation reaction. After the hapten is coupled with a carrier protein, vomitoxin molecules can be fully exposed outside the carrier protein, the immunogenicity of the antigen is enhanced, and an antibody with high titer and high sensitivity can be generated after the hapten is immunized by a host animal. Results of the embodiment show that after the vomitoxin antigen is subjected to mouse injection immunization, the titer of the generated antibody is 1: 20000, and the half inhibitory concentration (IC50) is less than 50ppb / mL. When the obtained vomitoxin antigen and antibody are applied to rapid quantitative detection of light-activated chemiluminescence of vomitoxin, the detection limit, the false negative rate and other dimensions are qualified, the detection steps are simple, and the vomitoxin antigen and antibody have good industrial value.

The invention discloses a method for preparing a high-titer polyclonal antibody from a Zika virus strain, which comprises the following steps: selecting the Zika virus strain, culturing in cells, and purifying virus particles with high purity to prepare an immunogen; an optimized animal immunization strategy is adopted and comprises the steps of selecting a proper host animal, applying a potent immunologic adjuvant, namely a Freund's adjuvant, and designing a scientific immunization program, so that humoral immune response of the host is stimulated to the maximum extent; when the titer of the antibody reaches a peak value, collecting antiserum, synthesizing hybridoma cells through splenocytes, and obtaining the high-purity polyclonal antibody through a multi-step purification technology. According to the method, high-purity and non-chemically-inactivated whole virus particles which retain natural conformation are used as immunogens, a mode of primary immunization and multiple booster immunization is adopted, a Freund's adjuvant is combined, animals are continuously stimulated to generate high-titer antibodies, non-specific antibodies and impurities are effectively removed through multi-step purification, and the immunogen immunogen is obtained. The target antibody with high purity and high specificity is obtained.