115results about How to "Stable secretion" patented technology

The invention disclose preparation of hybridoma cell, a monoclonal antibody secreted by the hybridoma cell and application of the monoclonal antibody and belongs to the field of microbial engineering. During screening of fusion cells prepared with the hybridoma cell, an antigen conjugated with magnetic beads and capable of specifically combining with a nonspecific antibody generated in cell culture is utilized; therefore the inhibitory effect of the nonspecific antibody upon positive reactions during the screening of the fusion cells is eliminated, and preparation efficiency is effectively improved. The invention further discloses H-FABP (Heart fatty acid-binding protein) resistant hybridoma cell strain, a monoclonal antibody produced by the strain, and application of the monoclonal antibody. The strain is capable of stably and effectively secreting high-specific H-FABP resistant monoclonal antibodies; the antibodies are of high potency and are applicable to various branch fields of in vitro diagnosis, especially immunodiagnosis.

The invention provides a monoclonal antibody TBCD6 resistant to mycobacterium tuberculosis culture filter protein (CFP) 10. A preparation method of the monoclonal antibody is characterized by immunizing the mice with the specific antigenic peptide, collecting sensitized B lymphocyte of the mice to undergo fusion with the myeloma cells of the mice, judging positive clones by utilizing enzyme-linked immunosorbent assay (ELISA) and western blotting, building a hybridoma cell system TBCD6 which secretes the monoclonal antibody, amplifying, culturing and injecting the positive cells to the peritoneal cavities of the mice of the same strain, using the mice to induce ascites containing the antibody and obtaining the monoclonal antibody through collection and affinity purification of the antibody. The invention provides application of the monoclonal antibody to detection and quantitative analysis of the mycobacterium tuberculosis CFP 10.

The invention discloses an anti-mycoplasma bovis monoclonal antibody, a hybridoma cell strain secreting the monoclonal antibody and an application. When the monoclonal antibody reacts with different mycoplasma strain mycoproteins, Western Blotting results indicate that the monoclonal antibody 3G11 only performs specific immune reaction with a mycoplasma bovis isolate, and does not react with mycoplasma mycoides SC (MmmSC), mycoplasma mycoides LC (MmmLC), mycoplasma capricolum pneumonia (Mccp), M.agalactiae, and M.ovipneumoniase. Therefore, the anti-mycoplasma bovis monoclonal antibody has good specificity, can be used for diagnosing and testing mycoplasma bovis infection, and provides an effective approach to prevent and control the disease.

The invention relates to a formula of an anti-stress extruded floating feed for megalobrama amblycephala and a preparation method thereof, belonging to the field of fish feed and preparation thereof. The anti-stress extruded floating feed for the megalobrama amblycephala, which comprises the following components by weight percent: 10-16% of fish meal, 15-30% of extruded soybean, 10-20% of rapeseed meal, 4-10% of peanut meal, 6-12% of cottonseed meal, 8-12% of corn protein powder, 10-18% of wheat shorts, 1-5% of squid paste, 0.25-0.75% of 50% choline chloride, 0.1-0.5% of acanthopanax root, 0.01-0.1% of pilose Asiabell root, 0.01-0.05% of Chinese magnoliavine fruit, 0.02-0.08% of schefflera arboricola hayata, 0.1-1% of spine date seed, 0.01-0.08% of jatamans valeriana rhizome, 0.5-1.5% of vitamin premix, 0.5-1.5% of mineral premix, 2-5% of calcium hydrophosphate, 1-2% of fish oil and 0.5-1.5% of lecithin oil. The anti-stress extruded floating feed for the megalobrama amblycephala can improve the immunity and the oxidation resistance of the megalobrama amblycephala, improve the stress and the anti-disease capability, improve the palatability of the feed, promote the growth and improve the survival rate of the megalobrama amblycephala.

The present invention relates to a gamma-interferon sandwich ELISA detection method based on recombinant fusion antigen protein, further relates to a recombinant fusion antigen protein preparation method and a gamma-interferon monoclonal antibody preparation method, and belongs to the technical field of medical examination determination. According to the present invention, the recombinant fusion antigen protein is formed by linking secretory antigen protein MPB70, antigen protein CFP10 and antigen protein ESAT6 through peptide bonds, the gamma-interferon monoclonal antibody is prepared through immunization of animals, cell fusion, hybridoma cell screening, cloning and purification, and the recombinant fusion antigen protein and the gamma-interferon monoclonal antibody are adopted to establish a sandwich ELISA detection method; and the recombinant fusion antigen protein can rapidly stimulate a bovine blood sample to secret gamma-interferon, and the gamma-interferon monoclonal antibody can quickly and specifically identify bovine gamma-interferon, such that strong sensitivity and strong specificity are provided, and broad application prospects are provided in the field of buffalo tuberculosis diagnosis.

The invention discloses a hybridoma cell strain RSVN4C3 secreting an anti-respiratory syncytial virus monoclonal antibody. The hybridoma cell strain RSVN4C3 is preserved in China Center for Type Culture Collection (CCTCC), and the preservation number is CCTCC NO: C2020254. The anti-respiratory syncytial virus nucleocapsid protein monoclonal antibody secreted by the hybridoma cell strain RSVN4C3 is used for preparing a kit for detecting or diagnosing respiratory syncytial virus infection, and the kit can be an immune colloidal gold test strip or an ELISA kit. The hybridoma cell strain RSVN4C3 can stably secrete the anti-respiratory syncytial virus monoclonal antibody (MAb), the reactivity and specificity of the monoclonal antibody (MAb) are identified, and a foundation is laid for future diagnosis of RSV virus infection and research of an infection mechanism.

The present invention relates to one kind of and its preparation and use, and belongs to the field of biotechnology. The HIV-1 virus-like particle contains complete core protein gag, encoding enzyme protein pol and outer membrane protein env of HIV-1, and is nonreplication type. The present invention is superior in that stably expression cell line is established through cotransfection and monoclonal cell screening. The cell line can secrete HIV-1 virus-like particle stably and continuously, and the HIV-1 virus-like particle contains no virus nucleic acid and has high safety. The constituted VLP has reasonable assembling form and high similarity with natural virus in structure, and may be used as HIV-1 treating vaccine.

The invention provides a nursing sow feed which comprises following ingredients in parts by weight: 40-70 parts of corn, 10-30 parts of soybean meal, 0-15 parts of flour, 1-5 parts of fish meal, 1-3 parts of fish oil, 0.4-1.0 part of calcium hydrophosphate, 0.1-0.4 parts of lysine hydrochloride, 0.1-1 part of arginine, 0.1-0.5 parts of valine, 0.08-0.2 parts of choline chloride, 0.03-0.10 parts ofgrowth promotion element, 0.1-0.4 parts of table salt and 4 parts of premix. The invention provides a preparation method of the nursing sow feed at the same time. The nursing sow feed is balanced innutrition, high in palatability and easy to digest, and can improve immunity of a sow; the feed intake of the sow can be increased significantly when the feed is eaten; the weight of weaning litter ofsuckling piglets is increased; a death rate of the piglets is reduced; and the quantity of the weaned piglets is increased.

The invention provides a hybridoma cell strain, a canine parvovirus VP2 protein monoclonal antibody generated by the hybridoma cell strain and application, and belongs to the technical field of monoclonal antibody preparation. The hybridoma cell strain 5A92B12 provided by the invention can be stably passaged and can continuously and stably secrete the canine parvovirus VP2 protein monoclonal antibody; after being cultured to 10 generations, the hybridoma cell strain 5A92B12 can still grow well and is stable in passage, and the supernatant titer of a culture solution can still reach 1x10<6> or above; the monoclonal antibody generated by the hybridoma cell strain and canine parvovirus VP2 protein can be specifically recognized; and the hybridoma cell strain has the advantages of high specificity and high sensitivity, has the characteristic of inhibiting canine parvovirus replication, can be applied to the detection and treatment of canine parvovirus diseases, and has a good application prospect.

The invention provides an immunodetection technology, and concretely relates to a ractopamine semiantigen and an artificial antigen, preparation methods thereof, and a monoclonal antibody prepared through immunizing mice by using the artificial antigen. The monoclonal antibody is secreted by a hybridoma cell strain 12B2, and the hybridoma cell is preserved China Center for Type Culture Collection with the preservation number of (CCTCC NO):C2014127. The ractopamine monoclonal antibody provided by the invention has the advantages of high titer, high sensitivity and strong specificity, and can be applied in the detection of the residual of ractopamine medicines.

The invention provides a monoclonal antibody TBEF3 of tubercle bacillus-resistant secretory antigen target protein in early stage. The monoclonal antibody TBEF3 is prepared by the following steps: selecting a specific antigen peptide mouse; collecting sensitized mouse B lymphocyte and mouse myeloma cell to fuse; judging positive clones by using an enzyme linked immunosorbent assayelisa and an immunoblotting testing method; establishing a hybridoma cell line TBEF3 for secreting monoclonal antibody of tubercle bacillus-resistant secretory ESAT-6 in early stage; carrying out multiplication culturing on the positive cells and injecting to congenic mouse enterocoelia for induction of generation of ascitic fluid containing antibody; and carrying out collection and antibody affinity purification to obtain the monoclonal antibody TBEF3 of tubercle bacillus-resistant secretory antigen target protein in early stage. The invention also provides an application of the monoclonal antibody in testing the secretory antigen target protein of mycobacterium tuberculosis in early stage.

The invention relates to an anti-progesterone monoclonal antibody with high specificity and affinity, a hybridoma cell generating the monoclonal antibody and application thereof, and particularly application in detecting the progestogen level of a human body. According to the invention, a progesterone complete antigen immune animal is adopted, cell fusion is performed after an ideal titer is achieved, and then the cell single strain with higher specificity is screened and preserved; and the strain single strain can continuously secrete a monoclonal antibody to facilitate the detection of progesterone, thereby laying a foundation for the development of a kit for detecting progesterone.

The invention relates to a bordetella pertussis PT (pertussis toxin) antigen monoclonal antibody and application thereof, and belongs to the field of preparation of biological products. The bordetella pertussis PT antigen monoclonal antibody is secreted by hybridoma cell strains capable of stably secreting bordetella pertussis FT antigen monoclonal antibodies, and a preservation number of the hybridoma cell strains is CGMCC No.10588. The bordetella pertussis PT antigen monoclonal antibody and the application have the advantages that the monoclonal antibody is high in potency and good in specificity, and can be applied to monitoring the quality in bordetella pertussis vaccine production procedures, carrying out enzyme-linked immunosorbent assay on contents of PT components in finished vaccine and preparing bordetella pertussis PT antigen enzyme-linked immune monitoring reagents or enzyme-linked immune monitoring reagent kits.

The invention discloses a monoclonal antibody against an extracellular domain of a goose CD3 epsilon chain and an application of the monoclonal antibody in detection of goose CD3<+> T lymphocytes. Amplification is performed by adopting goose thymus cDNA as a template to obtain a goose CD3 epsilon gene, according to the characteristics of a GoCD3 epsilon extracellular domain gene, a pair of specific primers are designed, amplification is performed to obtain the goose CD3 epsilon extracellular domain gene (CD3 epsilon ex) and the CD3 epsilon ex is expressed by virtue of escherichia coli; the recombinant plasmid containing the CD3 epsilon ex is adopted as an immunogen to immunize BALB/c mice, and the recombinant protein rGoCD3 epsilon is used for booster immunization to obtain a hybridoma cell strain (3C11) capable of stably secreting the monoclonal antibody against the extracellular domain of the goose CD3 epsilon chain. The monoclonal antibody can react with the purified recombinant protein rGoCD3 epsilon and can have a specific reaction with separated goose peripheral blood lymphocytes. Therefore, a novel and effective technical means is provided for detecting the goose CD3<+> T lymphocytes.

The invention provides a novel Avermectin resistant monoclonal antibody, hybridomas cell line excreting it and its preparing process, which comprises synthesizing artificial antigen AVM-BSA, immuning BALB/c mouse for five times, carrying out cell fusion to mouse spleen cells and SP2/O marrow tumor cells, using PEG1450 as fusion agent to screen positive fused cells, subcloning five times with limited dilution method, obtaining hybridomas cell lines 2F2 specifically secreting AVM antibody, whose micro-organism Deposit number is CGMCC NO.1413, injecting BALB/c mouse's abdominal cavity with the hybridomas cell, gathering ascitic fluid, centrifuging, gathering supernatant fluid, purifying and identifying to obtain the Avermectin resistant monoclonal antibody.

The invention relates to an animal pancreatic extract and a medicinal purpose of the animal pancreatic extract. In the invention, the animal pancreatic extract for preventing and curing diabetes is an active substance solution which is extracted from the fresh pancreas of a young pig and has small molecular weight not more than 10000 Daltons, the extraction liquid is supernatant liquid, the pH value of which is 6.0-7.0, and each 1ml of the extraction liquid contains not less than 15mg of polypeptide and does not contain insulin. The animal pancreatic extract of the invention is an active polypeptide substance with small molecular weight, which is obtained by taking the organ pancreas which causes the diabetes as a target spot, selecting the animal pancreas as a raw material and using the modern technology for extracting. The animal pancreatic extract of the invention is suitable for preventing and curing various types of diabetes, curing severe diabetes, preventing levis diabetes fromworsening, preventing organ posttransplantation diabetes, reducing adverse reaction of anti-rejection drugs, reversing or delaying the occurrence and the development of the severe diabetes, and preventing various complications of diabetes.

The invention discloses a hybridoma cell strain capable of secreting an African swine fever virus p34 protein monoclonal antibody, the monoclonal antibody and an application, the preservation number of the hybridoma cell strain is CCTCC NO: C202042, and the preparation method of the hybridoma cell strain comprises the following steps: 1) expressing to obtain an African swine fever virus p34 protein, 2) immunizing animals to obtain immunized spleen cells, 3) carrying out cell fusion and positive hybridoma cell strain screening, and 4) screening a positive hybridoma cell strain secreting a monoclonal antibody, wherein the p34 protein expressed by HEK293 cells transfected with CHO-S cells, SF9 insect cells and recombinant adenovirus vectors is adopted to screen positive hybridoma cell strains. The p34 antigen protein is prepared by adopting a mammalian cell CHO expression system, so that the folding and glycosylation of the protein are closer to those of natural protein, hybridoma cell strains are screened through the antigens prepared by the three expression systems, so that the obtained hybridoma cell strains can stably secrete monoclonal antibodies, and the specificity is stronger.

The invention discloses a monoclonal antibody and antibody combination for resisting a foot-and-mouth disease type O virus and application thereof in detection of antigens and antibodies of the virus, and provides a non-specific monoclonal antibody 4H9anti and antibody combination for the foot-and-mouth disease type O virus. The antibody combination comprises specific monoclonal antibodies 6D6anti and 1F2anti, a non-specific monoclonal antibody 2H10anti and the non-specific monoclonal antibody 4H9anti for the foot-and-mouth disease type O virus. The monoclonal antibody and antibody combination can realize detection of antigens and antibodies of the foot-and-mouth disease type O virus, can quickly detect multiple type O virus strains, have favorable broad-spectrum activity in detection of the foot-and-mouth disease type O virus, have the characteristics of high specificity, high sensitivity and high detection speed, and can be used for production of foot-and-mouth disease type O virus vaccines, quality control of related products and evaluation of immune effects of the vaccines.

The invention discloses a hybridoma cell strain, a getah virus distinguishing monoclonal antibody secreted from the hybridoma cell strain, and an application. The hybridoma cell strain is a K3A6B6 hybridoma cell strain secreting the monoclonal antibody for distinguishing getah virus E2 protein. The monoclonal antibody secreted from the hybridoma cell strain can be specially bound with target antigen in high titer, and the monoclonal antibody is prepared into a detection clinical sample for detecting a reagent kit. Specificity is high, sensitivity is high, and stability is good.

The invention provides an anti-MG7-Ag monoclonal antibody and an application thereof. The monoclonal antibody is an anti-MG7-Ag monoclonal antibody MGd1-Ab. The invention also provides a hybridoma cell line secreting the anti-MG7-Ag monoclonal antibody MGd1-Ab. The invention also provides a kit comprising the anti-MG7-Ag monoclonal antibody MGd1-Ab, and an application of the kit for detecting gastric cancer antigen MG7-Ag and an application of the kit for detecting expression level of the gastric cancer antigen MG7-Ag in a clinical tissue sample. The anti-MG7-Ag monoclonal antibody provided bythe invention has uniform texture and high specificity. The detection kit of the present invention can regulate sensitivity and detection range according to applications, thereby detecting expressionlevel of the gastric cancer antigen MG7-Ag in high sensitivity.

The invention relates to the biological engineering field, in particular to hybridoma of a DON (Deoxynivalenol) monoclonal antibody as well as a preparation method and application thereof. Through cell fusion, an immunizing antigen and a detection antigen adopt DON-BSA (Bovine Serum Albumin) to obtain and store hybridoma secreting the DON monoclonal antibody with high efficiency, and the antibody has high sensitivity, strong specificity and appetency with DON and low cross reaction with other toxins. The invention lays the foundation for researching DON test reagents.

The invention relates to a stem cell microsphere gel compound for subcutaneous injection and a preparation method and application of the stem cell microsphere gel compound. The preparation method comprises the following specific steps: 1, separating umbilical cord mesenchymal stem cells; 2, preparing stem cell-alginate microspheres; 3, preparing hydrogel; 4, mixing the stem cell-alginate microspheres with the hydrogel, wherein the dispersion concentration of the stem cells in the hydrogen is 105-106 per mL; and 5, subcutaneously injecting the stem cell microsphere gel compound to a wound in atissue to repair tissue injury. A high polymer material such as hyaluronic acid and stem cells are embedded and combined through microspheres, the microspheres are dispersed into gel to form a biological scaffold material with a function of fixing cell factors released from stem cells in an oriented manner, targeted repair and therapy effects of stem cells can be improved remarkably, a physical barrier is formed by gel so that stem cell tumor formation risks are avoided, and by the structures of the microspheres, the cell factors released from the stem cells have the characteristic of controllability in slow releasing, and thus, the stem cell microsphere gel compound conforms to development tendency of safe drug administration and accurate drug administration, and has high clinical application value.

The invention discloses encoded recombinant urate oxidase protein. A nucleotide sequence of the encoded recombinant urate oxidase protein is shown as SEQ ID NO:1. The nucleotide sequence of the encoded recombinant urate oxidase protein is connected with a phoA promoter and a periplasmic secretion signal peptide StII sequence and is transferred into escherichia coli to construct UOX escherichia coli engineering bacteria capable of being efficiently and stably secreted and expressed; then a low-phosphate culture medium is used for culturing; after fermentation is finished, centrifuging is carried out to remove supernatant of fermentation liquid, so as to obtain thalli; a TSE solution is used for extracting to obtain the recombinant urate oxidase protein. After purification is carried out, the urate oxidase protein is detected and has relatively good biological activity.

The invention discloses a hybridoma cell line of Escherichia-coli O157:H7 resistant monoclonal antibody. By adoption of the hybridoma cell line, the problem that the monoclonal antibody is not easily obtained; by passage for many years and times, the monoclonal antibody can be stably secreted, and the secreted monoclonal antibody is applied to a liquid-phase chip and a colloidal gold method to detect Escherichia-coli O157:H7 in foods, and has the advantages of good sensitivity, specificity, stability and repeatability and the like. Pathogens also can be detected under the condition of low concentration, so that the detection sensitivity and the detection rate are further increased and the leaking detection and the false detection are prevented strictly.

The invention relates to the technical field of biology, and discloses a hybridoma cell strain, a monoclonal antibody generated by the hybridoma cell strain and application. The hybridoma cell strainhas the preservation number being CGMCC NO.14290. The monoclonal antibody generated by the hybridoma cell strain can recognize the subgroup A avian leukosis virus gp85 envelope protein and can be usedfor developing subgroup A avian leukosis diagnosis and treatment reagents or medicine; a material basis is provided for subgroup A avian leukosis clinical differential diagnosis and laboratory research.