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公开(公告)号:US10766949B2
公开(公告)日:2020-09-08
申请号:US16540326
申请日:2019-08-14
发明人: Kun Li , Zengjun Lu , Zaixin Liu , Yimei Cao , Huifang Bao , Yingli Chen , Sheng Wang , Pinghua Li , Pu Sun , Xingwen Bai , Dong Li
IPC分类号: C07K16/10
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 method includes the following steps: 1) conducting the immunization on the cattle; 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; 4) acquiring constant region sequences of the heavy and light chains; 5) preparing a full-length heavy chain vector; 6) co-transfecting a cell, taking the supernatant of cell culture, and purifying. The method 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.
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2.
公开(公告)号:US20200055925A1
公开(公告)日:2020-02-20
申请号:US16540326
申请日:2019-08-14
发明人: Kun Li , Zengjun Lu , Zaixin Liu , Yimei Cao , Huifang Bao , Yingli Chen , Sheng Wang , Pinghua Li , Pu Sun , Xingwen Bai , Dong Li
IPC分类号: C07K16/10
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.
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