Gupta K C, Agha R, Santos E, Brodeur B R
Ryerson Polytechnical Institute, Toronto, Canada.
Hum Antibodies Hybridomas. 1992 Jan;3(1):25-31.
Human tonsillar lymphocytes immunized in vitro with diphtheria toxoid were fused with the heteromyeloma cell line SP2/HPT. Hybrids were selected on hypoxanthine-thymidine aminopterin (HAT) medium and assayed for antibody production both by ELISA and Western blotting. Four stable hybridoma cell lines producing 3-7 micrograms/mL of human IgG1 antibodies were isolated. In Western blotting studies these antibodies show binding to the B fragment of diphtheria toxin. Cells from the subclone P2-13/G35 were injected i.p. in pristane-primed nude mice, and ascites fluid was collected after 4-6 weeks. The monoclonal antibody neutralized the cytotoxicity of diphtheria toxin in the Vero cell assay system. This human monoclonal antibody may have a potential in the immunotherapy of diphtheric disease.
用白喉类毒素体外免疫的人扁桃体淋巴细胞与异骨髓瘤细胞系SP2/HPT融合。在次黄嘌呤-胸腺嘧啶核苷-氨基蝶呤（HAT）培养基上选择杂交瘤，并通过酶联免疫吸附测定（ELISA）和蛋白质印迹法检测抗体产生。分离出四个稳定的杂交瘤细胞系，它们产生浓度为3 - 7微克/毫升的人IgG1抗体。在蛋白质印迹研究中，这些抗体显示与白喉毒素的B片段结合。将亚克隆P2 - 13/G35的细胞腹腔注射到用 pristane 预处理的裸鼠体内，4 - 6周后收集腹水。该单克隆抗体在Vero细胞测定系统中中和了白喉毒素的细胞毒性。这种人单克隆抗体可能在白喉疾病的免疫治疗中具有潜力。
