论文部分内容阅读
以分泌抗恶性疟红内期抗体的小鼠杂交瘤细胞系94D1诱生的腹水,通过Protein A-Sepharose CL-4B亲和层析法纯化IgG,并以此作为免疫原皮下多点免疫Wistar大鼠。加强免疫后3天取脾与小鼠SP~2/0瘤细胞进行异种间的细胞融合,经间接ELISA法检测筛选和5次克隆化,最终得到一株生长稳定、连续分泌特异抗体的异种杂交瘤细胞系41RF5。经过多次冷冻和复苏以及体外连续传代培养16个月,其稳定性与小鼠×小鼠杂交瘤细胞相近。选择8株单抗和SP2/0IgG作为包被抗原,用间接ELISA法对41RF5特异性进行鉴定.结果显示,41RF5单抗只与94D1呈阳性反应,与其余各种包被抗原均呈阴性反应,表明41RF5具有良好的抗94D1单抗独特型决定簇的特异性。41RF5培养上清中特异的抗体效价为1:1280。
Ascites induced by mouse hybridoma cell line 94D1 secreting anti-Plasmodium macaque antibody was purified by Protein A-Sepharose CL-4B affinity chromatography and used as immunogen to immunize Wistar mouse. Three days after booster immunization, the spleen and the mouse SP2 / 0 tumor cells were harvested for cell fusion of different species. After screening and five clones by indirect ELISA, a stable and continuously secreting specific antibody was obtained. Tumor cell line 41RF5. After several freezing and resuscitation and continuous subculture in vitro for 16 months, its stability and mouse × mouse hybridoma cells similar. Eight monoclonal antibodies and SP2 / 0 IgG were chosen as coating antigen, and the specificity of 41RF5 was identified by indirect ELISA.The results showed that 41RF5 McAb was only positive with 94D1, and negative with all other coating antigens, This indicates that 41RF5 has a good specificity against idiotypic determinants of 94D1 mAb. The specific antibody titer in 41RF5 culture supernatant was 1: 1280.