Xu Yanmei, Yang Zhenggen, Liu Shili, Huang Huihui, Dong Wenqi
School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou 510000, China.
Guangzhou Koncen Bio Science Co., Ltd, Guangzhou 510000, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2020 Jul;36(7):640-644.
Objective To prepare the high-affinity and high-specificity mouse anti-human IgE monoclonal antibody (mAb) as a candidate immunosorbent. Methods BALB/c mice were immunized using recombinant antigen IgECepsilon2-4. Coated ELISA plate with IgECepsilon2-4 was used to screen positive cell lines by indirect ELISA, then coated ELISA plate with natural IgE, IgG, IgM, IgA, IgD to detect the affinity and specificity of serum-free culture supernatant of positive hybridoma cells with natural IgE. The cell line stably secreting specific anti-IgE monoclonal antibodies was expanded and inoculated into the abdominal cavity of BALB/c mice to prepare mAbs. The secreted mAbs were identified by ELISA kit followed by the identification of mAb subtypes. Results The 29 positive hybridoma cells were obtained after five cell fusions, of which 11 strains had strong affinity with natural IgE and 2 strains did not cross-react with other immunoglobulins 3E9 and 7B4. Conclusion The study successfully prepared mAbs against human IgE with high titer, affinity and specificity.
目的 制备高亲和力、高特异性的小鼠抗人IgE单克隆抗体（mAb）作为候选免疫吸附剂。方法 用重组抗原IgECε2-4免疫BALB/c小鼠。用IgECε2-4包被酶联免疫吸附测定（ELISA）板，通过间接ELISA筛选阳性细胞系，然后用天然IgE、IgG、IgM、IgA、IgD包被ELISA板，检测阳性杂交瘤细胞无血清培养上清与天然IgE的亲和力和特异性。将稳定分泌特异性抗IgE单克隆抗体的细胞系扩大培养并接种到BALB/c小鼠腹腔制备mAb。用ELISA试剂盒鉴定分泌的mAb，随后鉴定mAb亚型。结果 经过5次细胞融合获得29株阳性杂交瘤细胞，其中11株与天然IgE有强亲和力，2株3E9和7B4不与其他免疫球蛋白发生交叉反应。结论 本研究成功制备了高滴度、高亲和力和高特异性的抗人IgE单克隆抗体。
