2 results about "Brucella antibody" patented technology

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.