7 results about "Brucella antibody" patented technology

The application belongs to the technical field of biological pharmacy, and particularly relates to a bovine-derived mouse anti-Brucella antibody and a preparation method and application thereof. A bovine-derived mouse anti-Brucella antibody is prepared by synthesizing a tandem epitope protein of Brucella OMP25 proteins of bovine species, sheep testis species and canine species, immunizing mice, preparing a monoclonal antibody, cloning CDR fragments in Hv and Lv regions of the antibody to corresponding positions of bovine-derived IgG, and then performing overall mutation modification on the recombinant IgG to perform bovine-derived modification. The bovine-derived mouse anti-Brucella antibody preparation has good immunogenicity and protection effect, and can block and inhibit the propagation and spread of Brucella of bovine species, canine species and sheep testis species in an animal body.

The invention relates to the technical field of biological detection, and particularly discloses a preparation method and application of a Brucella antibody immunochromatography detection test strip based on colloidal carbon. The Brucella antibody immunochromatography detection test strip provided by the invention comprises a PVC (Polyvinyl Chloride) bottom plate, a sample pad, a combination pad, a nitrocellulose membrane and a water absorption pad are arranged on the PVC bottom plate; the binding pad contains lipopolysaccharide of colloidal carbon labeled brucella, the nitrocellulose membrane is provided with brucella lipopolysaccharide and a goat anti-mouse igG antibody which are respectively used as a detection line (T) and a quality control line (C), and the colloidal carbon technology-based immunochromatography test paper is established by detecting different specimens and comparing with other detection methods. The test strip prepared by the invention has the advantages of no subjective influence of a detector, simplicity and convenience in operation, high detection sensitivity, strong specificity and the like.

The application relates to the technical field of biological detection, and particularly discloses a bovine brucella antibody immunochromatographic detection test strip as well as a preparation method and application thereof. The bovine brucella antibody immunochromatographic detection test strip provided by the application comprises a PVC bottom plate, a sample pad, a combination pad, a nitrocellulose membrane and a water absorption pad are arranged on the PVC bottom plate, the combination pad contains quantum dot microsphere labeled mouse anti-bovine IgG monoclonal antibody, the nitrocellulose membrane is provided with a detection T line and a quality control C line, the detection T line is coated with a brucella LPS antigen, and the quality control C line is coated with a goat anti-mouse IgG polyclonal antibody. The bovine brucella antibody immunochromatographic detection test strip provided by the application has the advantages of being not influenced by subjective factors of a tester, simple and convenient operation, high detection sensitivity and specificity and the like.

The application belongs to the field of biotechnology, and provides a nano antibody against Brucella Omp25 protein and application thereof in competitive ELISA detection. The amino acid sequence of the nano antibody is shown as SEQ ID NO. 3. The application immunizes a llama with prokaryotic expressed Brucella Omp25 protein as an immunogen, extracts total RNA of peripheral blood lymphocytes, performs reverse transcription and two rounds of nested PCR to amplify a VHH fragment, performs enzyme digestion and connection on the VHH fragment and a pComb3Xss vector, and transforms the VHH fragment into TG1 competent cells. Through multiple rounds of panning by using phage display technology, the nano antibody against Brucella Omp25 protein is screened. The competitive ELISA detection method established by using the nano antibody can effectively detect the Brucella antibody level in sheep serum, has low detection limit and high sensitivity; and the developed matching kit has high specificity and greatly shortens the detection time.

The application discloses a kind of detection anti-Brucella antibody multi-link B cell epitope recombinant protein and its application, it belongs to the field of biotechnology and immunodiagnosis technique, wherein, multi-link B cell epitope recombinant protein is formed by the B cell epitope of seven key outer membrane proteins Omp10, Omp16, Omp19, Omp25, Omp31, Omp2b, BP26 of Brucella, its amino acid sequence is as shown in sequence table SEQ ID NO:1.The multi-link B cell epitope recombinant protein has good antigenicity and reactivity, and the deer serum indirect ELISA detection system established with it as coating antigen shows excellent detection performance, has high sensitivity, strong specificity and good repeatability, and can accurately detect the level of anti-Brucella antibody in deer serum sample.

This invention discloses a method for preparing Brucella antibodies, relating to the field of Brucella antibody preparation technology. Based on one or more combinations of Brucella outer membrane proteins Omp10, Omp19, and BP26, a recombinant antigen is prepared, and the prokaryotic expression codon is optimized. Alpacas are immunized subcutaneously with the recombinant antigen and adjuvant at multiple sites, with booster injections every two weeks for three weeks, to isolate highly active lymphocytes. Total RNA is extracted from the lymphocytes, reverse transcribed to synthesize cDNA, and the VHH gene is specifically amplified by nested PCR to accurately obtain the antibody coding sequence. The VHH gene is cloned into a phage vector, and a high-capacity display library is constructed by electroporation. Solid-phase panning is performed using the recombinant antigen as a target, followed by sequencing to construct a prokaryotic expression vector. Recombinant nanobodies are induced to express, purified by affinity chromatography, and elution conditions are optimized to obtain high-purity, high-yield target proteins. The optimized prokaryotic expression and purification process results in nanobodies with high soluble expression rate, high purification yield, and stable activity, achieving large-scale, low-cost preparation.

The application belongs to the technical field of animal epidemic disease immunity detection, and particularly relates to a Brucella antibody detection test paper and application, which comprises a bottom plate, a chromatography membrane arranged on the bottom plate, one end of the chromatography membrane is provided with a sample processing layer, and the chromatography membrane is further provided with a quality control line and at least one detection line; the sample processing layer comprises chicken IgY marked latex microspheres, Protein G protein marked latex microspheres or / and Fcα / μR receptor marked latex microspheres, and the detection line comprises Brucella LPS; or the sample processing layer comprises Brucella LPS marked latex microspheres and chicken IgY marked latex microspheres, and the detection line comprises Protein G protein and Fcα / μR receptor; the application significantly improves the sensitivity of early diagnosis or infection period discrimination, and is good in specificity, repeatability and accuracy.