抗旋毛虫单克隆抗体杂交瘤细胞系的建立

An Establishment of Monoclonal Antibody Hybrid Cell Line of Antitrichinella Spiralis

用旋毛虫感染性的幼虫和排泄分泌抗原免疫BALB/C纯系小鼠,取其脾细胞和SP_2/O—Ag14骨髓瘤细胞,在聚乙二醇(PEG)作用下融合,用间接酶联免疫吸附试验(ELISA)检测,阳性孔用有限稀释法经3～4次克隆化筛选培养,获得了D_4G_6和E_2B_(10)二个杂交瘤细胞系.经3～6个月的传代培养、冻存和接种小鼠诱生腹水,杂交瘤细胞生长良好,保持了稳定分泌抗体的能力.鼠性腹水和血清效价分别为6×10^(-3)和1×10^(-4).抗体免疫球蛋白用免疫琼脂双扩散试验测定均为IgG_1亚类.

BALB/C mouse was immunized with infective T.Spiralis larvae. Fusion of spleen cells and SP2/O-Agl4 were performed in the presence of polyethylene(PEG). Cultures were tested by indirect enzyme-linked immu-nosorbent assay (ELISA). Positive wells were cloned 3 - 4 times by limited dilution. Two hybrid cell lines-D4G6 and E2B10 were received and have the ability of secreting antibody steadily and grow very well, which has been proved by the experiment of 3-6 months, such as cell culturing, superfrozen storage and inducing abdominal liquid of inoculated mouse. The titer from mouse abdominal liquid is 6×10-3 and that from serum of mouse is 10-4. The antiboby both from D4G6 and E2B10 is IgG1 that was tested by agar bidirectional diffusion.