抗B血型物质单克隆抗体的研制及其免疫学特性的研究

PREPARATION AND IMMUNOLOGICAL CHARACTERIZATION OF MONOCLONAL ANTIBODIES TO B BLOOD GROUP SUBSTANCES

应用杂交瘤技术,分别以人B血型红细胞,B血型红细胞加纯化的B血型物质和B血型物质为抗原,采用不同免疫程序免疫BALB/C小鼠,从6次细胞融合中筛选建立起3株分泌高滴度,高特异性抗人B血型物质单克隆抗体的杂交瘤细胞株:3—3—D_9(IgG_1),3—5—D_(12)(IgG_1)和6—1—G_(11)(IgA),用Takatsy微量血凝滴定法测得它们的96孔板组织培养上清液的血凝滴度分别为4096,4096和16384。经过凝血特异性试验,凝血抑制试验以及抗体稀释试验等广泛免疫化学研究证明这些单克隆抗体仅对人B型血特异。杂交瘤细胞在液氮中贮存数月,分泌抗体能力不变。抗体经56℃保温30minA-20℃和37℃反复冻融试验凝血活性保持稳定。这些单克隆抗体为进一步研究B血型抗体结合部位结构的多样性提供了条件,并且使在临床血液分型试验时用单克隆抗体替代人多克隆抗血清成为可能。

Using hybridoma technic, we prepared a group of hybridomas by different immunization procedures with(?)B erythrocytes, B erythrocytes plus B blood group substances or B blood group substances alone as antigens ,respectively. Three of them: 3-3-D9(IgG1) , 3-5-D1 2 (IgG1) and 6-1-G11 ( IgA ) which secreted high level and high specific monoclonal anti-B blood group substances were established. The agglutination titers of their tissue culture supernatant were 4096, 4096 and 16 384 by Takatsy microtitration assays. It was demonstrated that they were only specific to B blood group by hemagglutination specificity tests, hemagglutination inhibition assays and antibody dilution tests. They will be very useful in blood grouping to replace the human poly-clonal anti-B sera. Studies to determinate their combining site structures will be possible by these monoclonal antibodes.