36 results about "Freund adjuvant" patented technology

The invention provides a polyvalent inactivity vaccine for preventing and treating atrophic rhinitis of a swine and a preparation method thereof. The polyvalent inactivity vaccine contains inactivated Bordetella bronchiseptica, Pasteurella multocida A, Pasteurella multocida D and PMT (Pasteurella Multocida Toxin) anatoxin. The invention further provides a novel method for culturing and extractingPMT. Compared with the traditional atrophic rhinitis of the swine, the polyvalent inactivity vaccine for the atrophic rhinitis of the swine, provided by the invention, can be used for more generally and effectively treating and preventing the atrophic rhinitis of the swine by comprehensive antigen protection. Finally, in the polyvalent vaccine provided by the invention, the vaccine with a plurality of antigens in a reasonable proportion can be used for solving the problem that the plurality of the antigens interfere each other, thereby improving an immune effect. Furthermore, the inventor provides a water adjuvant by which defects such as incomplete absorption, large side reaction and the like after the traditional alumina gel adjuvant, Freund adjuvant and water-in-oil adjuvant are injected into the water adjuvant can be overcome.

The invention discloses a Bacillus amyloliquefaciens WH3 strain, and a preparation method and application thereof. The preparation method comprises the following steps: 1, separation and identification of bacteria: separating bacteria resistant to rape sclerotinia rot from rape seedlings, and carrying out 16SrDNA and morphological identification to determine that the WH3 strain obtained by separation is Bacillus amyloliquefaciens; 2, separation, purification and identification of an antifungal active substance Safenour: fermenting the WH3 strain, extracting the antifungal active substance, separating and purifying through a sephadex column, and carrying out MALDI-TOF (matrix-assisted laser desorption/ionization-time of flight ) mass spectrometry on the active antifungal substance to inferthat the substance is a ring type polypeptide; and 3, application of the strain in the preparation of vaccines and immunological adjuvants. After the Safenour used as the adjuvant is mixed with a protein antigen and mice are respectively immunized through oral administration and injection of the mixture, effective body fluid and cellular immune response can be activated, and a high-titer specificantibody can be detected in the blood serum. The Safenour has low production cost and high stability, does not need to be emulsified when mixed with the antigen, and has a better immunoenhancement effect in comparison with Freund adjuvants and cholera toxin B subunits.

The invention discloses an acute monocytic leukemia-associated antigen MLAA-34 epitope polypeptide and an application of the acute monocytic leukemia-associated antigen MLAA-34 epitope polypeptide, and belongs to the technical field of biology. The acute monocytic leukemia-associated antigen MLAA-34 epitope polypeptide disclosed by the invention can induce specific CTL, and simultaneously can express tumor cells of MLAA-34 and HLA-A*0201, and T2 cells of loaded peptide in vitro to generate the determined specific killing effect; the epitope polypeptide CP701 is applied to preparation of a polypeptide vaccine aiming at acute monocytic leukemia; and the polypeptide vaccine composed of an MLAA-34 epitope peptide CP701 (236ILDRHNFAI244), a T auxiliary epitope and an incomplete Freund adjuvant can inhibit hu-PBL-SCID amplification of tumors in mice loading human monocytic leukemia cells THP-1, prolongs the survival life, and can induce the specific CTL killing activity and NK cytotoxic activity in vivo.

The invention relates to an amyloid protein intra-membrane segment for treating Alzheimer disease and an application thereof, in particular to the application of the amyloid protein intra-membrane segment in the preparation of a vaccine medicament for treating Alzheimer disease. The problem of treating Alzheimer disease can be efficiently solved. The technical scheme provided by the invention is to choose an intra-membrane segment (IF-A beta segment), which is called as AIIGLMVGGVVIA for short, from amino acid sequences of amyloid proteins (A beta42) of 42 amino acids. In a use process, after the synthesized IF-A beta and a Freund adjuvant are mixed and injected to an immunized AD (Alzheimer Disease) model mouse, the A beta42 plaques can be efficiently eliminated, the cognitive function of the AD mouse can be improved and the defect of the sub-acute meningitis caused by using A beta42 as an immunogen can be overcome, thereby developing a therapeutic medicament for efficiently treating AD without side effect.

The invention discloses a metformin monoclonal antibody hybridoma cell line and application thereof, and belongs to the field of food safety immunoassay. The prepared metformin monoclonal antibody hybridoma cell line Tcf 1A6 is deposited in the general microbiology center of the china microbial species collection management committee, the deposit date is March 7, 2019, and the preservation numberis CGMCC No. 17399. According to the metformin monoclonal antibody hybridoma cell line and application thereof, the complete antigen of the metformin is mixed and emulsified with an equivalent amountof Freund adjuvant, and BALB/c mice is immunized through multi-point subcutaneous injections of neck and back parts to screen hybrid cells after fusion of two kinds of cells; and then cells are screened through indirect competitive enzyme-linked immunosorbent assay and subcloned for three times, and the monoclonal antibody hybridoma cell line Tcf 1A6 is finally obtained. The provided monoclonal antibody secreted by the cell line Tcf 1A6 has better specificity and detection sensitivity to MET(the IC<50> value is 1ng/mL), the residual amount of MET in chicken, chicken liver, and feed can be detected, raw materials are provided for the immunoassay of the MET residues in food, and the practical applicational value is obtained.

The invention relates to a Listeria monocytogenes monoclonal antibody hybridoma cell strain and an application thereof and belongs to the technical field of food safety immunological detection. According to the invention, a complete antigen of a specific polypeptide of Listeria monocytogenes is uniformly mixed with an equal volume of Freund adjuvant (Sigma), BALB/c mice are immunized by subcutaneous injection, after the BALB/c mice are immunized for four times, spleen cells of immunized mice are fused with mice myeloma cells of immunized mice by virtue of a PEG method, an indirect ELISA screening and three times of subcloning are carried out to obtain a monoclonal cell strain A. The monoclonal antibody secreted by virtue of the monoclonal cell strain A has cross reaction with different Listeria monocytogenes culture supernatants and no cross reaction with listeria except Listeria monocytogenes, E. coliO 157, Salmonella and Campylobacter jejuni and thus the Listeria monocytogenes monoclonal antibody hybridoma cell strain can be used in the specific detection of Listeria monocytogenes in a food.

The invention relates to an application of krait venom antibacterial peptide in the preparation of a medicine for resisting inflammatory factor and belongs to the field of biomedical technology. A lot of researches show that the krait antibacterial peptide has an obvious effect of inhibiting inflammatory factor secretion induced by lipopolysaccharide in vitro and inhibits inflammatory factor secretion through inhibiting MAPK signal channels; the krait venom antibacterial peptide has a remarkable scavenging activity of DPPH and 2,2-azine-bis(3-ethylbenzothiazole-6-sulfoacid)diammonium salt (ABTS+)radical in vitro and has an obvious effect of inhibiting mice inflammation induced by a complete Freund adjuvant in vivo; and the krait venom antibacterial peptide can be applied in the preparation of a medicine for resisting inflammatory factor.

The invention relates to an immunolatex microsphere for detecting a cowpea trypsin inhibitor (CpTI) and a preparation method thereof. The immunolatex microsphere is coated with a CpTI-resistant protein polyclonal antibody and has the particle size of 200nm. The preparation method of the immunolatex microsphere comprises the following steps of: designing a full-length CpTI gene sequence; inserting a CpTI gene fragment into a glutathione-S-transferase (GST) gene fusion expression vector through restriction enzyme digestion to construct a bacteria fusion expression vector; purifying by using an affinity chromatographic column to obtain GST-CpTI fusion protein; fusing a GST-CpTI fusion protein antigen into a Freund adjuvant to immunize a mouse, and separating and purifying to obtain an anti-mouse CpTI polyclonal antibody; and coupling the antibody with the activated microsphere by a chemical coupling method to obtain the immunolatex microsphere for detecting the CpTI protein. The invention has the advantages that: the preparation method is high in sensitivity and repeatability and easy to operate; and the prepared immunolatex microsphere has small and uniform particle size, the lowest detectable limit of 0.2mu g/ml of the CpTI fusion protein, and the related coefficient R2 of 0.67.

The invention relates to the technical field of medicines, in particular to an application of an xanthiazone compound in preparing a drug for preventing or treating rheumatoid arthritis. A formula ofthe xanthiazone compound is shown as follows as shown in the specification. The compound significantly inhibits NO generation for RAW264.7 inflammatory cells and fibroblast-like synoviocytes stimulated by LPS (lipopolysaccharide), and has an obvious inhibiting effect on mouse ear swelling caused by dimethylbenzene and the rheumatoid arthritis induced by collagen, and an obvious swelling diminishing and anti-inflammatory effect on rat toe swelling due to inflammation caused by a Freund adjuvant; and therefore, the compound can be used for preventing and treating the rheumatoid arthritis.

The invention belongs to the technical field of biology, relates to a bone inflammatory pain model, in particular to application of a complete Freund adjuvant (CFA) to the preparation of a tibia inflammatory pain model. Tests on the tibia histopathology and behavioral indicators such as a heat pain threshold, a mechanical pain threshold and the like of the bone inflammatory pain model prepared by the CFA prove that: the inflammation of an experimental model is obvious, and the level of the behavioral indicators is low. A valuable bone inflammatory pain contrast model is provided for the research of bone cancer pain, particularly the mechanism research of cancer pain; a valuable experimental model is provided for the research of pain caused by clinical osteomyelitis; meanwhile, the type of a chronic pain experimental animal model is enriched, and the prepared tibia inflammatory pain model has important academic research value and application prospects.

The invention relates to a preparation method which comprises the following steps: taking a BALB/C mouse, extracting eye balls or sampling blood from an orbitall vein, carrying out EDTA-Na2 anticoagulation, separating a blood platelet in a gradient centrifugation manner and washing, and adjusting the concentration into 1 to 2*10/L; taking the suspension liquid of the blood platelet to be respectively mixed with equivalent complete freund adjuvant and incomplete freund adjuvant to form a milk white oil pocket water-shaped admixture which is used as antigen liquid; taking the antigen liquid containing the complete freund adjuvant to be respectively injected into the rear foot palm, the back and the groin of a guinea pig for four times, wherein each injection contains at least 5 points and the injection amount of each point is 100 microlitres; taking the whole blood of the guinea pig at the sixth week and taking blood serum for standby after the whole blood is centrifuged; taking the antiserum from -20 DEG C and placing the antiserum after being melted completely into water bath of 56 DEG C for 30 min; injecting the antiserum to the abdominal cavity of the BALB/C mouse on the first, the third, the fifth, the seventh, the ninth, the eleventh and the thirteen days and taking out on the fifteenth days, wherein the injection amount of each time is 100 microlitres. The invention solves the problem of shorter maintenance time of the blood platelet. The stable and controllable animal model of the idiopathic thrombocytopenic purpura of the mouse can be prepared.

The invention discloses application of a porphyrin pigment serving as immunologic adjuvant and further discloses application of the porphyrin pigment serving as a vaccine. The porphyrin pigment is a biological pigment, tissues outside an animal cardiovascular system can be combined with a pathogeny recognition receptor TLR4, and tissue damage and danger signals are simulated to build efficient stimulation to an immune system. The porphyrin pigment is also an ingredient of an animal and a human body, internal metabolic pathways are clear, uncertainty in the safety aspect does not exist, and the porphyrin pigment is suitable for preparation of the immunologic adjuvant and the vaccine for the animal, especially for human. The porphyrin pigment serving as the immunologic adjuvant can remarkably strengthen the humoral immunity effect of antigen or the vaccine and can reach the level of complete Freund adjuvant.

The invention discloses a method for gradient production of goat anti-Cys-C multiple resistant serum by means of amount of antigens. The method comprises the following steps: preparation of an immunogen emulsion: mixing and emulsifying 0.1ml of serum containing 4mg of the Cys-C antigen, 1.9ml of sterile PBS and 2ml of freund adjuvant to obtain an immunogen emulsion; first injection for goal immunity: injecting in two regions of each groin of front and rear groins of a goat, totaling 12 injection points; carrying out subcutaneous injection: performing average injecting at two injection points on two sides of the neck; injecting for the second time every 20 days, injecting for 3-8 doses every 10 days sequentially, and sampling blood from the artery of the immune goat every 4 days; putting the blood in a water tank for 1 hour at 40 DEG C; and after serum is fully separated out, centrifugalizing and precipitating to separate antiserum. The Cys-C antigen is subcutaneously injected to the groins of the goat for booster immunization for many times, so that the generated high-efficient goat anti-Cys-C antibody valence can reach over 1:16.

The invention discloses a pentachlorophenol monoclonal antibody hybridoma cell strain 2C3 and application thereof, and belongs to the field of food safety immunological detection. The pentachlorophenol monoclonal antibody hybridoma cell strain 2C3 is collected in the China General Microbiological Culture Collection Center (CGMCC) with a collection number CGMCC No.10871. A preparation method of the hybridoma cell strain 2C3 comprises the following steps: mixing a pentachlorophenol complete antigen and an equivalent Freund's adjuvant, and emulsifying the mixture, and immunizing a BALB/c mouse through back subcutaneous injection, wherein a complete Freund's adjuvant is used in a first immunizing process, and an incomplete Freund adjuvant is used afterwards; fusing spleen cells of the efficient and low-cost IC50 (Half Maximal Inhibitory Concentration) mouse with myeloma cells of the mouse through a PEG (Polyethylene Glycol) method, and performing indirect-competitive ELISA (Enzyme-Linked Immuno Sorbent Assay) screening and three times of sub-cloning to obtain a hybridoma cell strain 2C3. A monoclonal antibody secreted by the cell stain has high specificity and detection sensitivity (the IC50 value is 0.2mumg/L) specific to pentachlorophenol, and can be used for detecting residual pentachlorophenol in food safety.

The invention belongs to the technical field of biology, and discloses an acupuncture serum exosome and a preparation method and application thereof. The preparation method comprises the steps that S1, a rheumatoid arthritis model is established, specifically, a complete Freund adjuvant is injected to the sole of the right posterior foot of a rat to cause inflammation, after 24 hours, interventional electro-acupuncture treatment is conducted on the rat, the sole pain threshold of the rat is measured by using a full-automatic thermal radiation stimulator, and whether the rheumatoid arthritis model is successfully molded or not according to a pain threshold measurement result is judged; S2, serum extraction is conducted, specifically, abdominal aorta blood sample collection is conducted on the successfully molded rat, the blood sample is allowed to stand for 2 hours, centrifuging is conducted for 10 minutes in a centrifugal machine with the temperature of 4 DEG C and the speed of 2000rpm, after centrifugation, supernate is taken as serum and subpackaged in EP tubes, 500 microliters of serum is subpackaged in each EP tube, and all the EP tubes are put into a refrigerator at the temperature of -80 DEG C to be cryopreserved for use; and S3, exosome extraction is conducted, specifically, exosome is extracted from the serum extracted in the step S2 to obtain the acupuncture-like serum exosome capable of being used for treating rheumatoid arthritis.

The invention relates to an application of natural antibacterial peptide QHA in preparation of an immunologic adjuvant. The immunologic adjuvant is used for improving the immune titer of an antibody.In addition, the QHA and a Freund adjuvant have a synergistic effect when being combined for use. The invention discloses the novel application of the natural antibacterial peptide QHA. The natural antibacterial peptide QHA can be used for preparing the immunologic adjuvant for improving the immune titer of the antibody. The QHA as immunologic adjuvant for improving the immune titer of the antibody has wide application prospects in the fields of preparation of antibodies, vaccines, immunotherapy drugs, tumor vaccines or immune activators and the like.

The invention belongs to biological pharmaceutical field. It relates to a contraception vaccine cross linked with the molecule adjuvant, in particular to a hCGª‰-C3d3 confluence protein contraception vaccine cross linked with the molecule adjuvant C3d3. The invention selects eucaryon to express carrier phCMV1, constructs separately of the hCGª‰-C3d3 confluence protein with six histidine purification signals and eucaryon expressed plasmid phCMV1-6His-hCGª‰-C3d3 and phCMV1-6His-hCGª‰ of the hCGª‰protein, in vitro expresses, identifies and purifies hCGª‰-C3d3 confluence protein and hCGª‰protein, applies freunds adjuvant to immune mouse in different species. It is proved that hCGª‰-C3d3 confluence protein vaccine has comparative strong anti generating potent function by detection of the antibody level, biological activity of antiserum and neutralization of the hCG and excretion level of the cell factor by the spleen cells.