Klock G, Wisnewski A V, el-Bassiouni E A, Ramadan M I, Gessner P, Zimmermann U, Kresina T F
Department of Medicine, Miriam Hospital, Brown University, Providence.
Hybridoma. 1992 Aug;11(4):469-81. doi: 10.1089/hyb.1992.11.469.
Human monoclonal antibodies which bind Schistosoma mansoni worm and egg antigens were identified and characterized from hybridomas generated using the hypo-osmolar electrofusion technique of somatic cell fusion. Splenocytes from S. mansoni infected individuals were mitogen-activated in vitro and subsequently fused by electrofusion. The greatest number of HAT resistant hybridomas per helical fusion chamber was obtained with unfrozen splenocytes cultured for 4-6 days after introduction of mitogen. Hybridomas secreting IgG antibodies recognizing parasite antigens were identified by ELISA. Twenty-one cloned cell lines secreting IgG antibody were maintained for at least 6 months. Characterization of antigen reactivity by Western blot analysis of nien cloned cell lines revealed antibodies which bound stage specific parasitic antigens. The data show that the technique of hypo-osmolar electrofusion produces stable, antibody producing hybridomas. The human monoclonal antibodies screened represent candidate molecules useful in the investigations of the human pathogen S. mansoni.
利用低渗电融合体细胞融合技术从杂交瘤中鉴定并表征了与曼氏血吸虫虫体和虫卵抗原结合的人单克隆抗体。来自曼氏血吸虫感染个体的脾细胞在体外经丝裂原激活，随后通过电融合进行融合。在引入丝裂原后培养4 - 6天的未冻存脾细胞中，每个螺旋融合室获得的抗HAT杂交瘤数量最多。通过ELISA鉴定分泌识别寄生虫抗原的IgG抗体的杂交瘤。21个分泌IgG抗体的克隆细胞系维持了至少6个月。通过对11个克隆细胞系进行蛋白质印迹分析对抗原反应性进行表征，结果显示这些抗体与阶段特异性寄生虫抗原结合。数据表明，低渗电融合技术可产生稳定的、产生抗体的杂交瘤。筛选出的人单克隆抗体代表了可用于研究人类病原体曼氏血吸虫的候选分子。
