基于杂交瘤技术的BTN2A1单克隆抗体制备及其特异性研究OA

Aim To explore the preparation method of mono-clonal antibodies targeting BTN2A1 and develop detection tools,providing experimental evidence for optimizing immuno-therapy strategies.Methods High-affinity BTN2A1 monoclo-nal antibodies were screened using hybridoma technology;anti-body specificity for native conformational antigens was validated by flow cytometry;epitope mapping was performed via competi-tive ELISA;feasibility of a dual-antibody sandwich detection system was assessed based on antibody pairing experiments.Re-sults Fifteen high-affinity BTN2A1 monoclonal antibodies were successfully obtained(minimum half-maximal effective concentration:0.002 862 g·L-1).Flow cytometry confirmed that all antibodies specifically recognized cell-surface native BTN2A1 in BTN2A1-overexpressing CHO cells and Daudi tumor cell lines with>95%binding positivity.Epitope analysis indicated all antibodies targeted the same epitope.Conclu-sions This study successfully generates a panel of high-affinity BTN2A1 monoclonal antibodies,providing essential tools for functional research.However,since all antibodies recognize an identical epitope,establishing a dual-antibody sandwich detec-tion system is unfeasible.Further development of antibody com-binations targeting distinct epitopes is thus required,laying the foundation for optimizing BTN2A1 immunoassays and therapeu-tic antibody development.