摘要
用Protein A-Sepharose亲和层析法提纯McAb-IgG,皮下多点和腹腔免疫雄性Wistar大鼠,间隔两周共3次,融合前3天采用腹腔、尾静脉和脾内注射等三种加强免疫方法,以间接ELISA法和抗体竞争ELISA法筛选Anti-Id阳性克隆。结果显示:融合前大鼠血清抗小鼠IgG或抗免疫原McAb-IgG滴度,静脉组和脾内组比腹腔组高一个滴度;Anti-Id阳性克隆出现频率,腹腔组为0.9％和0.3％,静脉组为11.0％和13.3％,脾内组为16.4％和12.9％,静脉组和脾内组明显高于腹腔组(P<0.01);上述三种加强免疫方法对抗小鼠IgG阳性克隆出现频率无明显影响(P>0.05);与腹腔加强免疫组相比,静脉和脾内加强免疫组也可提高细胞融合效果(P<0.01).本实验结果提示,腹腔和皮下多点多次长程基础免疫结合静脉或脾内加强免疫是制备Anti-Id McAb比较好的免疫方案。
Anti-idiotypic antibodies are powerful reagents for the study of immunoregu-lation, and have potential interest as vaccines against tumors and infectious diseases. Three immunization, strategies for the production of rat monoclonal anti-idiotope antibodies have been compared in this paper. Male Wistar rats were immunized i.p. and at multiple subcutaneous sites with 750μg of purified monoclonal antibody against Plasmodium falciparum for three times and subsequently boosted by (1) intraperitoneal injection with 750μg of the immunogen, (2) intravenous inoculation with 400μg of the IgG, and (3) intrasplenic immunization with 200μg of the idiotype.With the intraperitoneal boost method, the frequency of hybrids with anti-idiotope activity was 0.3-0.9% with 62.8-85.2% of the seeded wells containing hybrids In the intravenous boost group, the percentage of hybrids demonstrating anti-idiotope activity increased to 11.0-13.3% with 80.2-97.9%of the hybrid efficiency. When immunized by the intrasplenic boost route, the frequency of anti-idiotope hybrids generated rose to 12.9-16.4% with 82.3-96.6% of the hybrid efficiency. There was no obvious effect of the boost immunizing methods on the generation of rat monoclonal anti-mouse IgG antibodies. These results indicated that the multiple-site immunization followed by intraveneous or intrasplenic boost injection was an appropriate immunizing method for the production of monoclonal anti-idiotope antibodies.
出处
《第一军医大学学报》
CSCD
1992年第3期193-198,共6页
Journal of First Military Medical University
基金
国家自然科学基金资助项目(No3880715)
关键词
抗独特型抗体
免疫
恶性疟
anti-idiotope
Plasmodium falciparum: heterohybridoma: monoclonal antibody
