摘要
采用不连续蔗糖密度梯度离心法纯化的鸡传染性法氏囊病病毒弱毒Gt株,并与Tj强毒株共同免疫BALB/c小鼠。利用淋巴细胞杂交瘤技术,取其脾细胞与SP2/0骨髓瘤细胞进行融合,经过3次筛选克隆,获得8株具有分泌抗IBDV抗体的杂交瘤细胞系,并对8株单抗进行了分子生物学及免疫学方面的鉴定。间接ELISA显示获得的8株单抗只与IBDV有特异性的反应,Western-blotting表明,部分单抗能够特异地与Sf9细胞表达的VP2蛋白反应;中和试验表明8株单抗均具有中和活性,而且有3株中和效价在26以上。研究推测,8株单抗中有2株针对的表位为构象依赖性,其余6株单抗所针对的为线性中和性表位。
The infectious bursal disease virus （IBDV） of Gt strain was purified by discontinuous gradient ultracentrifugation. After subcloning for three times, 8 strain hybridoma cells secreting monoclonal antibody（McAb） were established by the fusion of mouse myeloma cells SP2/0 and spleen cells from BALB/c mice immunized with purified IBDV Gt and Tj strain, and identification by immunologic and molecular biology characteristics. The McAb obtained from indirect ELISA had special reaction only with IBDV. Western blot indicated that partial McAb had specific reaction with the VP2 which was expressed on Sf9 cells. Neutralization test indicated that all of them have positive neutralization and potency of three McAb＇s over 26. Investigation was presumed that six of them were linear to IBDV antigen epitope, the others were conformational dependent form.
出处
《新疆农业大学学报》
CAS
2006年第2期17-21,共5页
Journal of Xinjiang Agricultural University
基金
国家重点基础研究发展计划(973项目)(2005CB523202)
关键词
传染性法氏囊病病毒
单克隆抗体
中和活性
infectious bursal diSease virus （IBDV）
monoclonal antibody （McAb）
neutrality activity
