946 results about "Artificial antigen" patented technology

The invention provides an artificial antigen presenting cell (AAPC) comprising a eukaryotic cell expressing an antigen presenting complex comprising a human leukocyte antigen (HLA) molecule of a single type, at least one exogenous accessory molecule and at least one exogenous T cell-specific epitope. Methods of use for activation of T lymphocytes are also provided.

T cell responses are often diminished in humans with a compromised immune system. We have developed a method to isolate, stimulate and expand naïve cytotoxic T lymphocyte precursors (CTLp) to antigen-specific effectors, capable of lysing tumor cells in vivo. This ex vivo protocol produces fully functional effectors. Artificial antigen presenting cells (AAPCs; Drosophila melanogaster) transfected with human HLA class I and defined accessory molecules, are used to stimulate CD8+ T cells from both normal donors and cancer patients. The class I molecules expressed to a high density on the surface of the Drosophila cells are empty, allowing for efficient loading of multiple peptides that results in the generation of polyclonal responses recognizing tumor cells endogenously expressing the specific peptides. The responses generated are robust, antigen-specific and reproducible if the peptide epitope is a defined immunogen. This artificial antigen expression system can be adapted to treat most cancers in a significant majority of the population.

The invention discloses an AFP[158-166] specific TCR gene, its transgenic T cell, and an in-vitro proliferation method and a use of the transgenic T cell. The AFP[158-166] specificity TCR gene is represented by SEQ ID NO.1 in a sequence table. The transgenic T cell of the AFP[158-166] specific TCR gene has a specific killing effect on AFP positive liver cancer cells. Artificial antigen presentation cells can present AFP[158-166] epitope peptides through the MHCI type molecule of the cell surface when the artificial antigen presentation cells are secreted by IL-15 to induce the AFP specific immune reaction, and the stimulation of the AFP[158-166] specific TCR gene transgenic T cell by the AFP[158-166] specific TCR gene can improve the proportion of the transgenic T cell, enhances the specificity and enhances the T cell activity and the antitumor capability in an auxiliary manner through the secretion of IL-15.

The present invention provides methods of preventing and / or treating cancers (including tumors). In one preferred embodiment, the invention is practiced to induce regression of an existing cancer or tumor and / or to prevent metastasis and / or to prevent growth of metastatic nodules. In other preferred embodiments, the invention may be used as a prophylaxis to prevent the development of primary cancers through a childhood or adult vaccination program against specific tumor antigens for cancers with high incidences. In an alternate preferred embodiment, the present invention provides methods of establishing an immune response against a universal artificial tumor antigen through a childhood or adult vaccine program, thus providing a long-term immune response that can be utilized at any point to treat any cancer which develops later in life. The present invention also provides cancer and tumor cells stably expressing an artificial antigen, preferably an artificial cell-surface antigen.

Carbon nanotube (CNT)-based compositions for activating cellular immune responses are provided. The CNTs function as high surface area scaffolds for the attachment of T cell ligands and / or antigens. The CNT compositions function as artificial antigen-presenting cells (aAPCs) or as modular vaccines. The disclosed CNT aAPCs are efficient at activating T cells and may be used to activate T cells ex vivo or in vivo for adoptive or active immunotherapy.

An artificial antigen presenting cell includes a liposome having at least one recombinant soluble MHC-peptide complex incorporated therein. The artificial antigen presenting cell may also include at least one additional signal molecule incorporated therein for manipulating the intensity and quality of the immune response. The recombinant soluble MHC molecule is obtained by a method utilizing PCR amplification of gDNA or cDNA, and a tag is attached thereto for anchoring the recombinant soluble MHC molecule to the liposome.

The invention provides an in vitro amplification method for efficient and highly cytotoxic natural killer (NK) cells. Novel artificial antigen presenting cells 4-1BBL-mIL-21-aAPC, such as 4-1BBL-mIL-21-K562 cells and the like, are constructed through stably expressing 4-1BB ligands (4-1BBL) and membrane immobilized interleukin 21 (mIL-21) on the surfaces of cell membranes, and by using the novel artificial antigen presenting cells as feeder cells for amplification, the NK cells are directly amplified from peripheral blood lymphocytes. Flow cytometry, cytotoxicity test and the like suggest that the amplified cells NK have high purity and strong cytotoxicity and have obvious killing effect on tumor cells.

The invention discloses a method for in vitro induction of specific cytotoxic T lymphocytes of hepatitis B virus (HBV). The method comprises the following steps of: 1, preparing artificial antigen presenting cells (aAPC) capable of loading any polypeptides; 2, loading aAPC on epitope peptides; and 3, loading aAPC in vitro induction HBV antigen specific CTL of antigen peptides; measuring the number of obtained antigen specific CTL by a Tetramer decoration method, namely combining streptavidin marked by fluorescein and four MHC-peptide compounds marked by biotin to form the Tetramer, namely MHC-peptide tetramer; and after the MHC-peptide tetramer is combined with TCR on the antigen specific CTL, measuring by a flow cytometry. The method can be applied to the amplification of T lymphocytes resisting the HBV.

The invention relates to a colloidal gold test strip for semi-quantitative detection of progesterone and a preparation method thereof. The invention belongs to the technical field of dairy cow pregnancy diagnosis. Colloidal gold test strip for semi-quantitative detection of progesterone, the sample pad, the gold pad containing gold-labeled progesterone antibody complex, and the nitrocellulose membrane are pasted end to end on the bottom plate; there are detection lines and control lines on the nitrocellulose membrane . Preparation of test strips: preparation of colloidal gold: prepared from chloroauric acid and trisodium citrate; preparation of purified polyclonal antibody: preparation of artificial antigen of dairy cow progesterone by dicyclohexylcarbodiimide coupling method; artificial antigen of progesterone was then used Immune mice to prepare progesterone polyclonal antibody; prepare gold-labeled antibody: colloidal gold and antibody adsorption, centrifugal purification; colloidal gold test strip preparation: soak gold-labeled pad with Tween-containing PB solution, gold-labeled antibody perfusion glass fiber Membrane; nitrocellulose membrane Spray bovine progesterone artificial antigen and goat anti-mouse secondary antibody into 2 lines with a film dispenser. The invention is simple in operation, convenient and practical, efficient and fast, and easy to popularize.

The invention discloses an immunological method for quickly detecting heavy metal lead ions and a kit. The immunological method is an indirect enzyme-linked immunological technology, and comprises the following steps of: coating lead ions in a lead-free antigen combined sample through artificially synthesized antigens to form lead-containing antigens; reacting the lead-containing antigens and anti-lead monoclonal antibodies to form an antigen-lead-antibody structure chain; reacting the structure chain and enzyme labeled secondary antibodies to form an antigen-lead-antibody-secondary antibody structure chain; and performing color development reaction, and judging the lead ion content of the detected sample according to the color development degree. The kit for implementing the immunological method comprises an artificial antigen coated and closed enzyme-linked immunosorbent assay (ELISA) plate and the anti-lead monoclonal antibodies, wherein the artificial antigen means CHX-A'-DTPA-OVA; and the anti-lead monoclonal antibodies are generated by monoclonal cell strains with collection date of August 25th, 2010, collection number of CCTCC C201084 and name of 6H8 / 4F7. The indirect enzyme-linked immunological detection technology is applied in small molecular detection for the first time, and the detection effect is good.

A synthetic method of a general artificial antigen of phthalate plasticizers for immunodetection belongs to the field of bio-chemical engineering technology. The synthetic method of the present invention comprises the following steps of carrying out an esterification reaction between phthalic acid and 6-(fluorenyl methoxy carbonyl acyl-amino)-1-hexanol, removing fluorene methoxy carbonyl acyl protecting groups to form a hapten, and coupling with amino from carrier protein to obtain the general artificial antigen of phthalate plasticizers. Experimental results disclose that titer of antiserum obtained by immunizing animals by the antigen of the invention is up to 160000;the 50% inhibiting concentration IC50 for DBP, DEHP and DINP is less than 500 ng / ml; and the generated antibodies have good generality and high sensitivity. The antigen or antibodies of the invention can be used for the establishment of enzyme-linked immunosorbent assay and colloidal gold test paper rapid detection methods, thus can be used for rapid detection of residues of phthalic acid ester plasticizers in food, and have broad application prospects.

The invention discloses the production method and use for imidacloprid artificial hapten, artificial antigen and specific antibody, wherein the production method comprises, using imidacloprid (1-(6-chlorine-3-picolyl)-N-nitro-2-imidazoline imine) as raw material for reaction with 3-mercaptopropionic acid under alkaline condition, thus synthesizing hapten 1-(6-(2-carboxyethyl) sulfo-3-picolyl)-N-nitro-2-imidazoline imines (IM), then coupling with proteins through carbodiimide method and mixed anhydride method to prepare artificial antigens (immunogens and peridium antigens).

The invention discloses a preparing method for paclobutrazol monoclonal antibodies. The preparing method comprises the main steps as follows: (1) synthesizing artificial hapten paclobutrazol hemisuccinates by a microwave solvent free method; (2) performing coupling to obtain artificial antigens of the paclobutrazol by using the paclobutrazol hemisuccinates as raw materials; (3) performing the immune treatment on Balb / c mice by using the synthesized artificial antigens; (4) selecting the mice with optimal serum valence and optimal specificity, and mixing spleen cells and myeloma cells of the mice together in an external manner; (5) culturing and screening the fused cells by using the selective culture medium, and further cloning, propagating and storing by freezing; (7) injecting the expanded cell strains into the abdominal cavities of the mice to generate a lot of abdominal dropsy; (8) purifying the paclobutrazol-resistant monoclonal antibodies in the abdominal dropsy by a caprylic acid-saturated ammonium sulfate method. The prepared paclobutrazol-resistant monoclonal antibodies have high sensitivity and strong specificity, and can be further applied to the construction of technologies of immune sensors, colloidal gold immune chromatographic methods and the like.

The invention provides an artificial antigen presenting cell as well as a preparation method and an application thereof. Wherein the artificial antigen presenting cell expresses membrane proteins MICA, IL-12 and CD80, the artificial antigen presenting cell is used for culturing NK cells or peripheral blood mononuclear cells containing the NK cells, the number of the NK cells can be remarkably increased, the service life of the NK cells is prolonged, the activity of the NK cells is improved, and compared with an existing artificial antigen presenting cell for expressing MICA, the artificial antigen presenting cell has a stronger effect of amplifying and activating the NK cells.

The invention discloses a preparation method and application of a product obtained by coupling melamine with carrier protein, as well as a melamine antibody. The product obtained by coupling the carrier protein with the melamine is used as artificial antigen and applied to an immunological method for melamine detection. The preparation method comprises the following steps of immunizing animals with the coupled product so as to prepare an antibody used for melamine detection, fusing BALB / C mouse spleen cells immunized with the coupled product and SP2 / 0 mouse myeloma cells, obtaining hybridoma capable of stably transferring culture and secreting anti-melamine specific monoclonal antibodies by screening positive hybridoma and cloning cells, and preparing an ascites monoclonal antibody. The prepared monoclonal antibody is utilized to establish a direct competitive ELISA method having high specificity, sensitivity and accuracy to the melamine, as well as an immune colloidal gold test strip. The preparation method for the product obtained by coupling melamine with carrier protein, as well as the melamine antibody provides service for the rapid detection of melamine-type residue in foods.

The invention relates to an immunoaffinity chromatography-ultrahigh performance chromatography-mass spectrum combined rapid detection method, and especially relates to a novel technology of a detection method for perfluorinated compounds in dairy products or water bodies. The method comprises following steps: step one, synthesis of artificial antigens; step two, screening and identification of polyclonal antibodies; step three, development of an immunoaffinity column; step four, establishment and optimizing of serially-combined detection method of ultrahigh performance liquid chromatography and mass spectrum. The technology provides a rapid detection method for perfluorinated compound residuals in the environment, improves the detection sensitivity, and guarantees the detection accuracy. The method meets the requirements on detection of perfluorinated compound residuals, and is beneficial for carrying out detections of perfluorinated compounds in real samples in the environment.

A fluo-insect nitrile artificial antigen, its specific antibody there-from and direct or indirect competitive enzyme immune adsorptive determining reagent kit are disclosed. The molecular structural formula is (I), n=1-5, in the (II), n is semi-antigen, and is covalent coupled with protein in proportion of 5:1-100:1 mol, protein is bovine serum albumin or egg-white protein. It can be used to detect fluo-insect nitrile residues in samples.

The invention provides Ciprofloxacin hapten, artificial antigen and antibody and a preparation method and application thereof. Ciprofloxacin is taken as a raw material, and is reacted with aminobutyric or aminocaproic to generate the hapten containing 4 to 6 chiral carbon atoms; the hapten is coupled with protein to prepare the artificial antigen by an active ester method; and the artificial antigen is used to immunize a mouse, and the monoclonal antibody with high specificity on the Ciprofloxacin is prepared by adopting cell fusion technology. The monoclonal antibody is adopted to establish immunological detection for quickly detecting the Ciprofloxacin on site, and has realistic significance for quickly detecting the Ciprofloxacin in livestock, poultry and aquatic products.

The invention discloses a method for detecting T-2 toxin and a special reagent kit thereof. The invention provides a hybridoma cell line 3E-5A-7H-8B whose preservation number is CGMCC No. 3395. The invention also provides a monoclonal antibody which is secreted by the hybridoma cell line 3E-5A-7H-8B whose preservation number is CGMCC No. 3395. The reagent kit provided by the invention comprises the monoclonal antibody. In the invention, the structure of the T-2 toxin is reconstructed, a space arm is added, and artificial antigens are synthesized. The artificial antigens are used for immuning animals to obtain the hybridoma cell line, and the monoclonal antibody which is secreted by the hybridoma cell line has high specificity. The reagent kit of the invention has the characteristics of simpleness of operation, low manufacture cost, high specificity, high sensibility, high precision and the like, the reagent kit which can be used for field monitoring is suitable for screening a great number of samples and can play an important role in detecting the T-2 toxin.

A flu-insect nitrile artificial semi-antigen, its synthesis and specific antibody made from it are disclosed. The molecular structural formula of semi-antigen is (I); the molecular structural formula of specific antibody is (II). It can be used to inspect flu-insect residues in sample.

A process for preparing artificial antigen, artificial hapten and specific antibody of chlorpyrifos is disclosed. A hapten O,O-diethyl-O[3,5-dichloro-6(2-carboxyethyl) thio-2- pyridyl] thionophosphate (AR) is prepared through reaction between O,O-diethyl-O(3,5,6-trichloro-2-pyridyl) thionophosphate and 3-hydroxypropionic acid. A hapten O-ethyl-O-[3,5,6-trichloron-(2-pyridyl)]-O- (3-carboxypropyl)thionophosphate (PO) is prepared from trichlorothion through 4-step reaction. An artificial antigen is prepared from said hapenjs and protein by coupling. A specific antibody is generated by immunizing animal with said antigen. It can be used to detect the residue of chlorpyrifos.

The invention discloses a process for preparing Quinclorac artificial semiantigen, artificial antigen, specific antibody and use thereof, wherein the preparation comprises, subjecting the dichloroquine (3,7-dichlorine-8-quinoline carboxylic acid) to sulfoxide chlorinated acylation, reacting with reanal and aminocaproic acid, thus obtaining semiantigen 4-(3,7-dichlorine-8-quinolineformyl) reanal or 6-(3,7-dichlorine-8-quinolineformyl) aminocaproic acid (QB or QC).

T cell responses are often diminished in humans with a compromised immune system. We have developed a method to isolate, stimulate and expand naïve cytotoxic T lymphocyte precursors (CTLp) to antigen-specific effectors, capable of lysing tumor cells in vivo. This ex vivo protocol produces fully functional effectors. Artificial antigen presenting cells (AAPCs; Drosophila melanogaster) transfected with human HLA class I and defined accessory molecules, are used to stimulate CD8+ T cells from both normal donors and cancer patients. The class I molecules expressed to a high density on the surface of the Drosophila cells are empty, allowing for efficient loading of multiple peptides that results in the generation of polyclonal responses recognizing tumor cells endogenously expressing the specific peptides. The responses generated are robust, antigen-specific and reproducible if the peptide epitope is a defined immunogen. This artificial antigen expression system can be adapted to treat most cancers in a significant majority of the population.

The invention provides a preparation method and application of a product obtained by coupling penicillin with carrier protein, as well as a beta-lactam type penicillin antibody. Animals are immunized with penicillin artificial antigen coupled in the invention so as to prepare the antibody which can be used for detecting beta-lactam type penicillin in foods. The preparation method comprises the following steps: immune BALB / C mouse spleen cells and SP2 / 0 mouse myeloma cells are fused; beta-lactam type antibiotics coupled with the carrier protein are used as coating antigen to screen positive hybridoma; hybridoma capable of stably transferring culture and secreting anti-beta-lactam type antibiotic antibodies through cell clones is obtained; and an ascites monoclonal antibody is prepared. The prepared monoclonal antibody is utilized to establish a direct competitive ELISA method having high specificity, sensitivity and accuracy to the beta-lactam type antibiotics, as well as an immune colloidal gold test strip. The preparation method for the product obtained by coupling penicillin with carrier protein, as well as the beta-lactam type antibiotic antibodies can serve the rapid detection of beta-lactam type antibiotic residue in foods.

The invention discloses a synthesis method of a specific salbutamol artificial antigen, belonging to the technical field of biological chemical engineering. The synthesis method disclosed by the invention comprises the following steps of: activating salbutamol through a formaldehyde solution, and connecting 6-aminocaproic acid in the ortho-position of a phenolic hydroxyl group to obtain a salbutamol hapten; and coupling a carboxyl group on the salbutamol hapten with an amino group on a carrier protein to obtain the salbutamol artificial antigen. The synthesis method disclosed by the invention can make up for the insufficiencies and defects of the existing salbutamol antigen synthesis technologies, the salbutamol artificial antigen with high specificity is obtained, the specificity of a produced antibody is high, and the sensitivity is high; and experimental results show that the antiserum titre of an animal immunized by using the salbutamol artificial antigen disclosed by the invention can achieve 80000, the detection limit is 0.5ng / mL, and the half-inhibitory concentration IC50 is 5ng / mL. The antigen or the antibody disclosed by the invention can be used for establishing an enzyme-linked immunosorbent analytical method and a colloidal gold test strip rapid assay method so as to rapidly detect the residues of the salbutamol in a food and further realize broad application prospects.

The invention relates to a analyzing method for hostathion direct competition enzyme immunity absorption analysis2 and its kit. Compose 4 hapten (the chemical formula)(n=1-5) and it covalency coupling symphysis with protein to be artificial antigen, use the artificial antigen immunity animal to prepare for antibody with idiosyncratic affinity to hostathion and use horseradish peroxide to mark the hapten. Use antibody to surround and cover the plate with microholes, add it into the mixture of the sample (or the marked standard sample of hostathion) and hapten marked by enzyme, hostathion, hapten marked by enzyme and the antibody on the surface of the polystyrene micro holes have competitive bond, then clear away the educt, add substrate and color-developing agent of enzyme the intensity of enzymatic colour reaction is inversely proportional to the content of hostathion in the sample (or the standard sample), in accordance with it, the hosathion direct competition enzyme immunity absorption analysis technique is invented. The technology can be applied to choose relative reagent and material in the kit, for preparing kit of immunity test, and rapid test for the polystyrene remained in agricultural products and environment.

The invention discloses a hybridoma cell strain secreting an anti-amantadine monoclonal antibody and application of the hybridoma cell strain. The name of the hybridoma cell strain is hybridoma cell strain 7G4.3, and the collection number is CCTCC No. C2014169. The hybridoma cell strain is obtained by immunizing a mouse with a self-designed amantadine artificial antigen and using spleen cells of the immunized mouse, the anti-amantadine monoclonal antibody secreted by the hybridoma cell strain has high titer and strong specificity, and can be used for fast and accurate immunodetection and immunoassay of amantadine.

The invention discloses an alternarin tenuazonic acid artificial antigen, a polyclonal antibody, a preparation method and application, and particularly discloses an artificial antigen obtained by coupling a compound (formula I) serving as a tenuazonic acid hapten, a hapten TAO, bovine serum albumin, hemocyanin and other macromolecular protein, a polyclonal antibody prepared from the artificial antigen and application of the artificial antigen and the polyclonal antibody to tenuazonic acid detection. TA (tenuazonic acid) can be detected by setting up an indirect competitive ELISA method, a theoretical basis is provided for a TA immunological detection method, a foundation is laid for further developing and detecting TA immunology fast detection products, and a new path is provided for setting up the TA immunological detection method, developing the TA immunological series products, and fast and efficiently monitoring TA contamination conditions.

The invention relates to a preparation method and use for 2, 4, 6-tri-chlorophenol artificial antigen. It is synthesized by hapten 2, 4, 6-trichlorobenzende oxygen acetic acid through activating fat or mixing acid anhydride. The antibody can be used to make 2, 4, 6-tri-chlorophenol antibody, set 2, 4, 6-tri-chlorophenol indirect competing fluorescence immunity method, or testing its residuum in the aquatic environment. The advantages of the invention are that the antibody has great practicability and good stability; the preparation method is easy and feasible; the cost is low; and it is easy to produce in industrial scale.

The invention discloses a 3-methylquinoxaline-2-carboxylic acid artificial antigen and an antibody obtained by the 3-methylquinoxaline-2-carboxylic acid artificial antigen. The invention provides a compound shown in the formula (I). The compound shown in the formula (I) is prepared by structural modification of 3-methylquinoxaline-2-carboxylic acid, retains a feature structure of 3-methylquinoxaline-2-carboxylic acid as much as possible and has active groups which can couple with a carrier protein. The 3-methylquinoxaline-2-carboxylic acid artificial antigen provided by the invention is a conjugate prepared by coupling of the compound shown in the formula (I) and a carrier protein. The 3-methylquinoxaline-2-carboxylic acid artificial antigen is used for animal immunization so that high-specificity monoclonal and polyclonal antibodies are obtained. A preparation method of the high-specificity monoclonal and polyclonal antibodies is simple and practicable. The 3-methylquinoxaline-2-carboxylic acid artificial antigen can be used for detection of olaquindox metabolites. The antibodies obtained by the 3-methylquinoxaline-2-carboxylic acid artificial antigen can be used for detection of olaquindox metabolites.