56 results about "Freunds Adjuvant" patented technology

The invention relates to a hybridoma cell strain secreting thiamethoxacin monoclonal antibody and application thereof, belonging to the field of food safety immunodetection. The accession number of the hybridoma cell strain is CGMCC No. 14699. According to the invention, a complete Freund's adjuvant is used for primary immunization of a BALB / c mouse, then an incomplete Freund's adjuvant is used for booster immunization three times, and a thiamethoxam complete antigen containing no adjuvant is used for impact immunization once, so the BALB / c mouse is immunized; and then the high-titer low-IC50spleen cells of the immunized mouse are fused with mouse myeloma cells by using a PEG method, and then the cell strain is obtained through indirect competitive ELISA screening and subcloning three times. The monoclonal antibody secreted by the cell strain has good specificity and detection sensitivity (with an IC50 value of 0.81 ng / mL) to thiamethoxam and can be used for detection of thiamethoxamresidues in food.

The invention discloses a hybridoma cell strain CBC secreting a monoclonal antibody of dicofol and application of the hybridoma cell strain CBC, and belongs to the field of food safety immunoassay. The cell strain CBC is preserved in the China General Microbiological Culture Collection Center, the address is Institute of Microbiology, Chinese Academy of Sciences, No.3, Beichen West Road, ChaoyangDistrict, Beijing, the preservation date is March 7, 2019, and the preservation number is CGMCC No.17398. Complete antigens of the dicofol are mixed and emulsified with equivalent freund adjuvants, and BALB / c mice are immunized by subcutaneous multipoint injection of neck and back through first immunization, multiple strengthening immunization and sprint immunization. Spleen cells of the mice withhigh titer and low IC50 are fused with mouse myeloma cells by a PEC method, selective culture is conducted, then cells are screened and sub-cloned for three times through indirect competitive enzyme-linked immunosorbent assay, finally the cell strain CBC is obtained, the monoclonal antibody secreted by the cell strain CBC has good detection sensitivity to the dicofol (value of the IC50 is 19.09 ng / mL), and can be used for the detection of dicofol residues in food.

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 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 discloses a preparation method of a porcine TLR4 polyclonal antibody. According to the preparation method, recombinant TLR4 protein (antigen) and freund's adjuvant are emulsified, then an immune animal obtains polyclonal antibody serum, wherein the recombinant TLR4 protein has the amino acid sequence as indicated in the sequence table SEQ. ID. No. 1. It is shown through tests that the antibody prepared through the method is good in reactivity and specificity; due to the fact that a TLR4 gene is a member of a significant pattern recognition receptor Toll-like receptor family targeted to innate immune response, the porcine TLR4 polyclonal antibody can be applied to detection of the porcine TLR4 protein and related research; a good foundation is laid for researching the interaction mechanism between porcine bacterial and viral infection diseases and TLR4 and the virus immunity escape mechanism, and a new target is provided for developing new generation vaccine and medicine. Meanwhile, a prokaryotic expression vector constructed through the method is high in recombinant protein expression efficiency, and the separation and purification method is simple and easy to operate.

The invention discloses an ethyl maltol monoclonal antibody hybridoma cell strain and application thereof, and belongs to the field of food safety immunodetection. The disclosed ethyl maltol monoclonal antibody hybridoma cell strain ABC01 is preserved in China General Microbiological Culture Collection Center (CGMCC) and is classified and named as a monoclonal cell strain on November 28, 2019, andthe preservation number is CGMCC No.19178. A complete antigen of ethyl maltol and an equivalent Freund's adjuvant are mixed and emulsified, and a BALB / c mouse is immunized through neck and back subcutaneous multipoint injection. Cells are screened with an indirect competitive enzyme-linked immunosorbent assay method and subcloned for three times, and finally, the monoclonal antibody hybridoma cell strain is obtained. The monoclonal antibody secreted by the cell strain has better specificity and detection sensitivity to ethyl maltol, can realize detection of the residual amount of ethyl maltolin grease, milk and other foods, provides a raw material for immunodetection of ethyl maltol residues in foods, and has practical application values.

The invention discloses a rabbit-derived anti-porcine parvovirus type 6 VP2 protein antibody and a preparation method thereof. The preparation method comprises the following steps: designing a pair of specific primers according to a PPV6 epidemic strain genome sequence published by GenBank, constructing a pET30a-PPV6-VP2 recombinant expression vector, carrying out IPTG induced expression, Ni-NTA resin affinity chromatography purification and urea dialysis renaturation at different gradients, and performing uniform mixing and emulsification with a Freund's adjuvant in a ratio of 1: 1; and preparing PPV6 recombinant VP2 protein immunogen. A New Zealand white rabbit is subjected to multi-point subcutaneous injection on the back, hyperimmune serum is prepared, and the antibody is purified by adopting a saturated (NH4)2SO4 salting-out method. The prepared rabbit-derived anti-porcine parvovirus type 6 VP2 protein antibody is has good purity without bacteria, mycoplasma or exogenous virus pollution, and has good specificity without other common swine pathogen antibodies except the porcine parvovirus type 6 VP2 protein antibody, and the characteristics sufficiently meet the requirement for positive serum for exogenous virus detection, so that an important significance on healthy development of the veterinary biological product industry is achieved.

A hybridoma cell strain secreting a nifursolol residue marker monoclonal antibody with CGMCC No. 14698 is prepared in the following way: BALB / c mice are subjected to the first immunization with a complete Freund's adjuvant, subjected to booster immunization with an incomplete Freund's adjuvant for four times, and subjected to rush immunization once with nifursolol residue marker complete antigen without a Freund's adjuvant so that the BALB / c mice are immunized; the spleen cells of the immunized mice with high titer and low IC50 were fused with mouse myeloma cells by a PEG method, and the fused cells are screened through indirect competitive ELISA and subcloned three times. The monoclonal antibody secreted by this cell strain has good specificity and detection sensitivity (IC50 value of 2 μg / L) to the nifursolol residue marker and can be used for residue detection of the nifursolol residual marker in food.

The invention provides a hybridoma cell strain E secreting an anti-glibenclamide monoclonal antibody and application thereof, and belongs to the field of food safety immunodetection. The hybridoma cell strain E secreting the anti-glibenclamide monoclonal antibody is deposited in CGMCC, the common microbiology center of the China Microbial Culture Collection Management Committee, and is classifiedas a monoclonal cell strain, the deposit date is May 24, 2018, and the deposit number is CGMCC No.17305. According to the hybridoma cell strain E, a complete antigen of glibenclamide and an equal amount of Freund's adjuvant are mixed and emulsified, and immunization BALB / c mice are subjected to subcutaneous multiple point injection atbacksand necks to finally obtain the hybridoma cell strain E secreting the anti-glibenclamide monoclonal antibody. The monoclonal antibody secreted by the cell strain has good specificity and detection sensitivity (IC50 value of 0.8 ng / mL) for the glibenclamide,can detect the residual amount of the glibenclamide in health foods, provides raw materials for the immunodetection of glibenclamide residues in the foods, and has a practical application value.

The invention discloses a forchlorfenuron monoclonal antibody hybridoma cell strain and an application thereof, and belongs to the technical field of food safety immunodetection. The preservation number of the hybridoma cell strain is CGMCC No. 19179. Forchlorfenuron hapten and complete antigen are firstly synthesized, mixing and emulsifying are performed by using a Freund's adjuvant, and injecting is performed to immunize a BALB / c mouse. High-titer and low-IC50 mouse splenocytes are screened, and are fused with mouse myeloma cells by a PEG method, a culture medium is selected, and hybrid cells obtained by fusing the two cells are screened. The cells are screened by an indirect competitive enzyme-linked immunosorbent assay and subcloning is performed for five times to finally obtain the monoclonal antibody hybridoma cell strain. The monoclonal antibody secreted by the cell strain has good detection sensitivity to the forchlorfenuron, can be used for preparing forchlorfenuron immunoassay kits and colloidal gold test strips, and is used for residue detection of the forchlorfenuron in foods.

The invention discloses a virginiamycin monoclonal antibody hybridoma cell strain YSL and application thereof, and belongs to the field of food safety immunodetection. The produced virginiamycin monoclonal antibody hybridoma cell strain YSL is preserved in China General Microbiological Culture Collection Center, the preservation date is March 7, 2019, and the preservation number is CGMCC No.17397.According to the virginiamycin monoclonal antibody hybridoma cell strain YSL and application thereof, by conducting mixing emulsification on a complete antigen of virginiamycin and the same amount ofFreund's adjuvant, conducting multi-point injection immunization on BALB / c mice, then screening out cells through an indirect competitive euzymelinked immunosorbent assay method and conducting subcloning for three times, the monoclonal antibody hybridoma cell strain YSL is obtained finally. A monoclonal antibody secreted by the provided cell strain YSL has good specificity and detection sensitivity (an IC50 value is 40 ng / mL) for VIR, the residual amounts of VIR in chicken, livers of chickens and feed of the chickens can be detected, and a raw material is provided for immunodetection of residual VIR in food, so that the virginiamycin monoclonal antibody hybridoma cell strain YSL has practical application values.

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 yersinia enterocolitica monoclonal antibody hybridoma cell strain and application thereof, and belongs to the field of food safety immunodetection. The yersinia enterocolitica monoclonal antibody hybridoma cell strain JSL3H3 is classified and named as a monoclonal cell strain, the preservation date is September 27, 2020, and the preservation number is CGMCC (China General Microbiological Culture Collection Center) No.20786. The monoclonal antibody secreted by the cell strain has good specificity and detection sensitivity for yersinia enterocolitica. The yersinia enterocolitica complete antigen is synthesized, the complete antigen and an equal amount of a freund's adjuvant are mixed and emulsified completely, a BALB / c mouse is immunized through back subcutaneous injection, and the hybridoma cell strain JSL 3H3 is obtained through screening of an indirect competitive enzyme-linked immunosorbent assay and three times of subcloning. The monoclonal antibody secreted by the cell strain has good specificity for the yersinia enterocolitica, and the results of the invention can be used for specific detection of yersinia enterocolitica in food, and has practical application value.

The invention relates to a lincosamides universal monoclonal antibody hybridoma cell strain and an application thereof, which belong to the technical field of food safety immunology detection. The lincosamides universal monoclonal antibody hybridoma cell strain employs a lincosamide substituted derivative as semiantigen, semiantigen is coupled with bovine serum albumin (BSA) through an activated ester method to obtain immunization antigen, the immunization antigen and a freund's adjuvant are uniformly mixed, a mixture is used for immunization BALB / c mice through hypodermic injection, clindamycin is coupled with albumin from chicken egg white (OVA) by employing a carbonyl diimidazole (CDI) method, and the material is used for detecting mice serum and cell supernatant as coating antigen, thespleen cell of the immunization mice is subjected to fusion with mice myeloma cells through a PEG method, and through indirect ELISA and indirect competition ELISA screening and three-time subclone,the group selected hybridoma cell strain can be obtained. The cell has good inhibition effect for clindamycin, lincomycin and pirlimycin, and can satisfy the requirement on lincosamides stumpy immunodetection products on the market.

The invention discloses a dihydropyridine monoclonal antibody hybridoma cell strain and application thereof, and belongs to the field of food safety immunodetection. The dihydropyridine monoclonal antibody hybridoma cell strain ABC02 is preserved in China General Microbiological Culture Collection Center (CGMCC), is classified and named as a monoclonal cell strain, and is preserved on November 28,2019 with the preservation number being CGMCC No. 19177. A dihydropyridine hapten is synthesized, then a dihydropyridine complete antigen is prepared, and is mixed with an equivalent Freund's adjuvant for thorough emulsification, a BALB / c mouse is immunized through subcutaneous injection on the back, fusion with myeloma cells is realized through a PEG method, and the hybridoma cell strain ABC02 is obtained through screening of indirect competitive enzyme-linked immunosorbent assay and three times of subcloning. The monoclonal antibody secreted by the cell strain has good specificity and detection sensitivity (IC50 is 0.35 ng / mL) to dihydropyridine, can be prepared into a kit for detecting dihydropyridine, and has practical application value.

The present invention discloses a tacrolimus monoclonal antibody hybridoma cell strain and an application thereof, and belongs to the technical field of immunodetection. The tacrolimus monoclonal antibody hybridoma cell strain ICP2B3 is classified and named as a monoclonal cell strain, the preservation date is November 28, 2019, and the preservation number is CGMCC No.19172. Hapten is synthesizedthrough tacrolimus oximation, and tacrolimus complete antigen is prepared and completely mixed with an equal amount of Freund's adjuvant for complete emulsifying. By immunizing BALB / c mice through back subcutaneous multipoint injection, high-titer and low-IC50 mouse spleen cells are selected to fuse with myeloma cells, screening is conducted through an indirect competitive enzyme-linked immunosorbent assay, subcloning for three times is also conducted, the monoclonal antibody secreted by the cell strain has relatively good specificity and detection sensitivity on tacrolimus (IC50 is 0.996 ng / mL) . The result can be used for establishing an immunodetection method for tacrolimus content in a clinical blood sample and has practical application value.

The invention discloses a hybridoma cell strain JS secreting an anti-phenolphthalein monoclonal antibody and application of the hybridoma cell strain JS, and belongs to the field of food safety immunodetection. The hybridoma cell strain JS secreting the anti-phenolphthalein monoclonal antibody is preserved in the China General Microbiological Culture Collection Center (CGMCC), the preservation date is March 7, 2019, and the preservation number is CGMCC No.17394. According to the hybridoma cell strain JS secreting the anti-phenolphthalein monoclonal antibody and the application of the hybridoma cell strain JS, a complete antigen of phenolphthalein and a Freund's adjuvant with the amount the same as that of the complete antigen are subjected to mixed emulsification, and a BALB / c mouse issubjected to subcutaneous multi-point injection at the nape part for being immunized. An indirect competitive euzymelinked immunosorbent assay is adopted for screening cells, subcloning is conductedthree times, and finally, the monoclonal antibody hybridoma cell strain is obtained. The monoclonal antibody secreted by the cell strain has good specificity and detection sensitivity (the IC50 valueis 15.4 ng / mL) for the phenolphthalein, the amount of phenolphthalein residuals in healthcare food can be detected, the raw material is provided for immunodetection for the phenolphthalein residuals in food, and the hybridoma cell strain JS secreting the anti-phenolphthalein monoclonal antibody has practical application value.

The invention discloses hybridoma cell lines capable of secreting diphenhydramine monoclonal antibodies and an application thereof, and belongs to the field of cosmetic safety immunodetection. The hybridoma cell lines SHS 5B11 capable of secreting diphenhydramine monoclonal antibodies are named as monoclonal cell lines by classification, and preserved on September 27, 2020 with the preservation number of CGMCCNo.20787. The monoclonal antibodies secreted by the cell lines have good specificity and detection sensitivity to diphenhydramine. According to the present invention, diphenhydramine complete antigens are synthesized, then subjected to mixed emulsification with equal amount of Freund's adjuvant, and afterwards subcutaneously injected to immune BALB / c mice through backs of the mice. The hybridoma cell lines SHS 5B11 are obtained through screening with indirect competitive enzyme-linked immunosorbent assay and subcloning three cloning. The monoclonal antibodies secreted by the cell lines have good detection sensitivity ( IC50 value of 2.93 ng / mL) to diphenhydramine, and can be used for detecting diphenhydramine residues in cosmetics.

The invention discloses a sirolimus monoclonal antibody hybridoma cell strain and application thereof, and belongs to the technical field of immunodetection. The invention discloses a sirolimus monoclonal antibody hybridoma cell strain ROX1C5, the strain is preserved in China General Microbiological Culture Collection Center (CGMCC) and is classified and named as the monoclonal cell strain, the preservation date is November 28, 2019, and the preservation number is CGMCC No.19171. Sirolimus hapten is synthesized through oximation reaction, sirolimus complete antigen is prepared, the sirolimus complete antigen and an equal amount of Freund's adjuvant are mixed and emulsified completely, and BALB / c mice are immunized through back subcutaneous multipoint injection; high-titer and low-IC50 mouse splenocytes are selected to be fused with myeloma cells, screening is conducted through an indirect competitive enzyme-linked immunosorbent assay, and subcloning is conducted three times so as to obtain the cell strain. The monoclonal antibody secreted by the cell strain has good specificity and detection sensitivity (IC50 is 3.90 ng / mL) on sirolimus, is used for establishing an immunodetectionmethod of sirolimus content, and has practical application value.

The invention discloses a hybridoma cell strain capable of secreting a modafinil monoclonal antibody and application of the hybridoma cell strain and belongs to the field of food safety immunodetection. Two modafinil haptens are provided, a modafinil complete antigen is prepared, the modafinil complete antigen and an equal amount of a Freund's adjuvant are mixed and emulsified completely, a mouse immunization experiment is carried out, high-titer and low-IC50 mouse splenocytes are taken and fused with myeloma cells through a PEG method, and the hybridoma cell strain is obtained through screening by an indirect competitive enzyme-linked immunosorbent assay and three times of subcloning. The monoclonal antibody secreted by the cell strain has relatively good specificity and detection sensitivity (IC50 is 0.54ng / mL) to modafinil. The result of the invention can be used for establishing an immunodetection method for modafinil content in human plasma, and thus, the hybridoma cell strain has a practical application value.

The present invention discloses a preparation method of an anti-RET mutant protein monoclonal antibody variable region sequence. Mutation is performed at the C634 position of a RET protein to synthesize a polypeptide sequence, and the polypeptide and a carrier protein KLH are respectively coupled and combined with freund's adjuvant for emulsification to prepare an emulsified antigen, mutants are separately used to construct expression sequences to a purple fluorescent protein mcherry amino terminus to be combined with an expression vector to prepare a detection agent, mice are immunized with the emulsified antigen and mouse myeloma SP2 / 0 cells and spleen cells of the immunized mice are subjected to cell fusion, and are cultured with a semi-solid culture medium and the detection agent, by observing formation of clone balls and color changes of the culture medium, positive clones are selected into new 6-well plates and subjected to expanded culture by a changed liquid culture medium, each cell line is then transferred to a shake flask for culture, a supernatant of the culture medium is collected, antibodies are purified, and the antibodies are sequenced to obtain the variable regionsequence.

The invention relates to a hybridoma cell line secreting acyclovir monoclonal antibody and a preparation method thereof, belonging to the field of food safety immunoassay. The preservation number of the hybridoma cell line is: CGMCC No.14695. BALB / c mice were first immunized with complete Freund's adjuvant, boosted with incomplete Freund's adjuvant, and adjuvant-free adjuvant BALB / c mice were immunized by shock immunization with Lowe's complete antigen; the immunized mouse splenocytes with high titer and low IC50 were fused with mouse myeloma cells by PEG method, and were screened by indirect competition ELISA and subcloned three times , the resulting cell lines. The monoclonal antibody secreted by this cell line has good specificity and detection sensitivity to acyclovir, and can be used for the detection of acyclovir residues in food.

The invention discloses a phosphorylated antigen polypeptide, a phosphorylated antibody and a preparation method for the phosphorylated antibody, and relates to the technical field of antibody preparation. The phosphorylated antigen polypeptide comprises the phosphorylated antibody which aims at human ERK (Extracellular regulated protein kinases) protein threonine 202 and tyrosine 204, and the amino acid sequence of the phosphorylated antibody is disclosed by SEQ ID NO.1, wherein Thr (threonine) and Try (tyrosine) are mutated into Asp (aspartic acid); and the amino acid sequence of the phosphorylated antibody which aims at YAP1 (Yes-associated protein 1) protein serine 127 is disclosed in SEQ ID NO:2, wherein Ser (serine) is mutated into Asp. A phosphorylated amino acid is mutated into theaspartic acid, the DNA (deoxyribonucleic acid) of a mutant antigen is constructed to a pGEX-6p expression carrier, escherichia coli is used for expressing a GST (glutathione S-transferase) fusion antigen, and after the purified antigen and Freund's adjuvant are mixed, an animal can be directly immunized. The method is simple, has good antigen stability, and is high in solubility and low in cost.

The invention relates to an immune checkpoint inhibitor related myocarditis mouse model and a construction method, and belongs to the technical field of medical animal models. The immune checkpoint inhibitor related myocarditis mouse model is constructed by subcutaneously injecting a complete Freund's adjuvant of a mouse cardiac troponin I peptide fragment into a mouse and injecting a PD-1 inhibitor into an intraperitoneal cavity of the mouse. According to the immune checkpoint inhibitor related myocarditis mouse model and the construction method, the success rate of modeling is effectively improved by determining the reasonable dosage and administration interval, the model construction method is simple and easy to implement and easy to popularize and apply, and the immune checkpoint inhibitor related myocarditis mouse model constructed by the invention provides an experimental object for the research of clinical immune checkpoint inhibitor related myocarditis pathogenesis and the screening of related myocarditis treatment drugs.

The invention discloses a hybridoma cell strain for secreting a nitrovin monoclonal antibody and application of the hybridoma cell strain, and belongs to the field of food safety immunodetection. The hybridoma cell strain FQJ 5G11 for secreting the nitrovin monoclonal antibody is preserved in the China General Microbiological Culture Collection Center (CGMCC), is classified and named as a monoclonal cell strain, and has a preservation date of September 27, 2020 and a preservation number of CGMCC No.20784. The method comprises the following steps of synthesizing a nitrovin hapten, preparing a nitrovin complete antigen, mixing and emulsifying the nitrovin hapten and the nitrovin complete antigen with an equivalent Freund's adjuvant completely, immunizing a BALB / c mouse through back subcutaneous injection, and performing screening by an indirect competitive enzyme-linked immunosorbent assay and three times of subcloning to obtain the hybridoma cell strain FQJ 5G11. The obtained nitrovin monoclonal antibody cell strain can be used for immunoassay detection, and has relatively good detection sensitivity and specificity for nitrovin.