目的：制备抗水通道蛋白4(aquaporin 4，AQP4)单克隆抗体，鉴定其免疫学性能为后期临床应用奠定基础。方法：合成AQP4优势抗原表位多肽，经过免疫小鼠、细胞融合及亚克隆筛选，制备可分泌高效价高亲和力的单克隆抗体细胞株，后期获取大量单克隆抗体通过酶联免疫吸附和免疫组织化学实验进行初步实验应用。结果：获得4株杂交瘤细胞株，均能稳定分泌高亲和力抗体，经酶联免疫吸附测定实验和免疫组织化学实验证实均能特异性识别人的AQP4蛋白。结论：成功制备了亲和力高、特异性强的抗人AQP4优势抗原表位的单克隆抗体，为小分子抗体以及抗体人源化制备奠定了基础，进而为治疗视神经脊髓炎相关疾病提供理论依据和技术支持。
Objective: To lay the foundation for clinical applications in the future, we prepare and identify the immunological properties of aquaporin 4 (AQP4) monoclonal antibodies. Methods: The dominant epitope polypeptides of AQP4 were synthesized and used to obtain the cell lines which secrete high tiler and high affinity monoclonal antibodies by immunizing BALB/c mouse cell fusion and screening of subclone. The large amount of monoclonal antibodies were obtained and used for practice via immunohistochemical staining and Enzyme-linked immunosorbent assay (ELISA). Results: Four hybridomas that can stably secret anti-AQP4 antibodies were obtained. ELISA, Western blot and Immunohistochemistry results suggested that the monoclonal antibodies specifically recognized human AQP4 protein.Conclusion: The anti-human AQP4 monoclonal antibody with high affinity and specificity was successfully generated, which thereby provides the foundation for the preparation of small molecule antibodies or humanized antibodies, and then lays a theoretical and technical basis for the treatment of neuromyelitis optica (NMO).
目的：制备抗水通道蛋白4(aquaporin 4，AQP4)单克隆抗体，鉴定其免疫学性能为后期临床应用奠定基础。方法：合成AQP4优势抗原表位多肽，经过免疫小鼠、细胞融合及亚克隆筛选，制备可分泌高效价高亲和力的单克隆抗体细胞株，后期获取大量单克隆抗体通过酶联免疫吸附和免疫组织化学实验进行初步实验应用。结果：获得4株杂交瘤细胞株，均能稳定分泌高亲和力抗体，经酶联免疫吸附测定实验和免疫组织化学实验证实均能特异性识别人的AQP4蛋白。结论：成功制备了亲和力高、特异性强的抗人AQP4优势抗原表位的单克隆抗体，为小分子抗体以及抗体人源化制备奠定了基础，进而为治疗视神经脊髓炎相关疾病提供理论依据和技术支持。
Objective: To lay the foundation for clinical applications in the future, we prepare and identify the immunological properties of aquaporin 4 (AQP4) monoclonal antibodies. Methods: The dominant epitope polypeptides of AQP4 were synthesized and used to obtain the cell lines which secrete high tiler and high affinity monoclonal antibodies by immunizing BALB/c mouse cell fusion and screening of subclone. The large amount of monoclonal antibodies were obtained and used for practice via immunohistochemical staining and Enzyme-linked immunosorbent assay (ELISA). Results: Four hybridomas that can stably secret anti-AQP4 antibodies were obtained. ELISA, Western blot and Immunohistochemistry results suggested that the monoclonal antibodies specifically recognized human AQP4 protein. Conclusion: The anti-human AQP4 monoclonal antibody with high affinity and specificity was successfully generated, which thereby provides the foundation for the preparation of small molecule antibodies or humanized antibodies, and then lays a theoretical and technical basis for the treatment of neuromyelitis optica (NMO).
