METHOD FOR PREPARING WHOLE BOVINE-DERIVED BROADLY NEUTRALIZING ANTIBODY AGAINST SEROTYPE O FOOT-AND-MOUTH DISEASE VIRUS

摘要

The present invention relates to a method for preparing a whole bovine-derived broadly neutralizing antibody against serotype O foot-and-mouth disease virus, and belongs to the technical field of antibody preparation. The preparation method of the present invention includes the following steps: 1) conducting the first, second and third immunization on the cattle by using the serotype O foot-and-mouth disease virus; 2) screening for the serotype O foot-and-mouth disease virus antigen-specific single B cells; 3) amplifying variable region genes of the heavy and light chains of the bovine antibody; 4) acquiring constant region sequences of the heavy and light chains of the bovine antibody; 5) preparing a full-length heavy chain vector of the whole bovine-derived monoclonal antibody and a full-length light chain vector of the whole bovine-derived monoclonal antibody; 6) co-transfecting a cell with the full-length heavy chain vector of the whole bovine-derived monoclonal antibody and the full-length light chain vector of the whole bovine-derived monoclonal antibody, taking the supernatant of cell culture, and purifying to obtain the whole bovine-derived broadly neutralizing antibody against the serotype O Foot-and-mouth disease virus. The preparation method of the present invention utilizes different foot-and-mouth disease virus strains to infect cattle and obtains a neutralizing antibody capable of neutralizing three topotypes of serotype O FMDVs by screening, and thus can obtain a whole bovine-derived broadly neutralizing monoclonal antibody.