摘要
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。
A hybridoma cell line secreting monoclonal antibody to idiotope of the monoclonal antibody 94Dl specific for asexual erythrocytic stages of Plasmodium falci-parum was established by fusion of SP2/0 mouse myeloma cells with spleen celle of Wistar rats immunized with monoclonal IgG of 94Dl purified from ascitic fluid of BALB/C mouse by affinity chromatography on Protein A-Sepharose CL-4 B. Specificity of the anti-idiotope antibody ( anti-Id ) , 41RF5, was was determined with indirect enzyme-linked immunosorbent assay ( ELISA ) . The results showed that 41RF5 reacted only with 94Dl IgG2b, not with normal mouse IgG and other seven mouse monoclonal antibodies-five specific for erythrocytic stages of P . falciparum, one for erythrocytic stages of P . inui, and one for Dengue fever virus type III, which indicates that 4lRF5 does recognize specifically the idioty-pic determinant of 94D1 monoclonal antibody. In 4lRF5 heterohybridoma culture supernatant, anti-Id titre measured by ELISA was above 1:1,280. Up to the present, the heterohybrid cell line has been cultured stably for 16 months.
出处
《第一军医大学学报》
CSCD
1989年第1期29-32,共4页
Journal of First Military Medical University
关键词
单克隆抗体
疟疾
杂交瘤
疟原虫
idiotype
monoclonal anti-idiotope antibody
mouse-rat heterohy-bridoma
Plasmodium falciparum
