55 results about "Freunds Adjuvant" patented technology

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 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 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 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 provides a hybridoma cell line secreting anti-bisamide compound monoclonal antibody and application thereof, belonging to the technical field of immunochemistry. The deposit number of the hybridoma cell line is: CGMCC No.22322. The present invention mixes and emulsifies the complete antigen with Freund's adjuvant, and subcutaneously immunizes mice to obtain high-titer, low-IC 50 Mouse splenocytes were fused with mouse myeloma cells by the PEG method, and the hybrid cells after the fusion of the two cells were screened out using selective medium; the cells were then screened by indirect competitive enzyme-linked immunosorbent method and subcloned three times, and finally obtained A monoclonal antibody hybridoma cell line. The monoclonal antibody secreted by this cell line has good detection sensitivity to cyclic bromantraniliprole, cyantraniliprole, chlorantraniliprole, and tetrachlorantraniliprole, and can be used for the detection of these four pesticide residues in food.

The invention relates to the application of a natural antibacterial peptide QHA in the preparation of an immune adjuvant, which is used to improve the immune titer of antibodies. In addition, the combination of QHA and Freund's adjuvant has a synergistic effect. The invention discloses a new application of the natural antibacterial peptide QHA, which can be used to prepare an immune adjuvant for improving the immune titer of antibodies. As an immune adjuvant for improving the immune titer of antibodies, QHA has broad application prospects in the fields of preparing antibodies, vaccines, immunotherapy drugs, tumor vaccines or immune activators.

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 invention discloses a novel application of brucella flagellin BME II 1112 in preparation of brucella subunit vaccines. The experimental results show that a BME II 1112 recombinant protein and a freund's adjuvant are subjected to combined immunization to be induced so as to produce a specific antibody, stimulate the cell immunity, and generate a better immune protecting effect for the brucella, thereby being capable of being utilized to prepare the brucella subunit vaccines. The application of brucella flagellin BME II 1112 in preparation of brucella subunit vaccines plays an important role in the epidemic prevention of the brucella, so that the application prospect is wide.

The invention relates to a hybridoma cell strain secreting meloxicam monoclonal antibody and a preparation method of the hybridoma cell strain and belongs to the field of food safety immunodetection.The hybridoma cell strain has a collection number of CGMCC No. 14700. The preparation method comprises the following steps: performing primary immunization on BALB / c mice with a complete Freund's adjuvant, performing booster immunization for three times by using an incomplete Freund's adjuvant, performing impact immunization once on an adjuvant-free meloxicam complete antigen, and enabling the BALB / c mice to be immunized; fusing high titer low-IC50 immunized mice spleen cells with mice myeloma cells by a PEG method, and performing indirect competitive ELISA (Enzyme-Linked Immuno Sorbent Assay)screening and three-times subcloning, thereby obtaining the cell strain. The monoclonal antibody secreted by the cell strain has excellent specificity and detection sensitivity (IC50 value of 0.1ng / mL) on meloxicam, and can be used for detecting residual meloxicam in foods.

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.