通过免疫β3Gn-T5 敲除小鼠生成并鉴定针对 GM3（NeuGc）神经节苷脂的 IgG 单克隆抗体。
Generation and characterization of a IgG monoclonal antibody specific for GM3 (NeuGc) ganglioside by immunizing β3Gn-T5 knockout mice.
机构信息
Department of Clinical Bio-Cell, 4th Hospital, Hebei Medical University, Shijiazhuang, 050000, China.
Department of Oncology, Hebei General Hospital, Shijiazhuang, 050000, China.
出版信息
Sci Rep. 2018 Feb 7;8(1):2561. doi: 10.1038/s41598-018-20951-8.
A murine monoclonal antibody (MAb-1) specific for GM3 has been generated by immunizing β3Gn-T5 knockout mice with purified GM3 ganglioside. The binding specificity of MAb-1 (IgG subclass) was established by an enzyme-linked immunosorbent assay (ELISA) and FACS and the antibody showed high binding specificity with GM3. Cell viability assay showed that MAb-1 significantly suppressed cell growth. Immunohistochemistry analysis revealed that MAb-1 was strongly expressed in human ovarian cancer tissues, whereas it was hardly expressed in normal tissues. Finally, antibody-dependent cellular cytotoxicity (ADCC) activities were determined by measuring lactate dehydrogenase (LDH) releasing assay and the results showed high ADCC activities in two representative ovarian cancer cell lines (OVHM and ID8). All of these data indicate that MAb-1 may be potentially used as a therapeutic antibody against ovarian cancers in clinical trials.
已通过用纯化的 GM3 神经节苷脂免疫β3Gn-T5 基因敲除小鼠，生成了一种针对 GM3 的鼠源单克隆抗体（MAb-1）。通过酶联免疫吸附试验（ELISA）和流式细胞术确定了 MAb-1（IgG 亚类）的结合特异性，并且该抗体与 GM3 表现出高度的结合特异性。细胞活力测定表明 MAb-1 可显著抑制细胞生长。免疫组织化学分析显示，MAb-1 在人卵巢癌组织中强烈表达，而在正常组织中几乎不表达。最后，通过测量乳酸脱氢酶（LDH）释放试验确定抗体依赖性细胞毒性（ADCC）活性，结果表明在两个代表性卵巢癌细胞系（OVHM 和 ID8）中具有高 ADCC 活性。所有这些数据表明，MAb-1 可能潜在地用作临床试验中针对卵巢癌的治疗性抗体。
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