摘要
目的:建立能稳定分泌抗呼吸道合胞病毒(RSV-long株,国际标准株)单克隆抗体的杂交瘤细胞株。方法:杂交瘤技术制备单抗,鉴定各项特性。结果:培育出稳定分泌抗RSV蛋白的单克隆抗体的杂交瘤细胞5株。杂交瘤细胞株染色体数目在89～104条之间,其分泌的抗体分别属于鼠IgG1、IgG2a、IgG2b亚类,各种腹水单抗的荧光效价在1∶4000～1∶16000之间。一种单抗识别RSV基质蛋白(M),两种单抗识别RSV融合蛋白(F),另两种单抗识别RSV核衣壳蛋白(N)。单抗中和效价最高达1∶64。相加指数证实5种单抗识别不相关的抗原表位。用硫氰酸铵洗脱法比较了五种单抗相对亲和力的大小,依次为:1#>2#>4#>3#>5#。所有杂交瘤细胞株,经连续3个月培养及冻存半年后复苏,细胞生长良好,检测上清,其分泌抗体效价稳定。结论:已获得抗呼吸道合胞病毒的单克隆抗体,为RSV感染的早期诊断及进一步研究奠定了基础。
Objective ：To establish the hybridoma cell lines stably secreting monoclonal antibodies against respiratory syncytial virus （RSV）. Methods：McAbs were prepared by hybridoma technique, and identified in terms of well-known criteria. Results：Five hybridoma cell lines secreting monoclonal antibody against RSV were established. The chromosome numbers of these cell lines were between 89 and 104. The McAbs were IgGl, IgG2a and IgG2b subclasses. The ascites titres of these McAbs were between 1 ： 4 000 and 1：16 000 by IFA. The highest titer was 1 ： 64 in the five McAbs. 1 # McAbs was against matrix protein of RSV, demonstrated with Western blot or immunoprecipitation, 2# and 4# of the McAbs were against fusion protein of RSV, 3# and 5# McAbs were against nucleocapsid-associated protein of RSV. ELISA additivity test proved the five kinds of McAbs recognized the unique antigen epitopes. The elution with thiocyanate showed that relative affinity of the five kinds of McAbs was as follows： 1# 〉 2# 〉4# 〉 3# 〉 5#. The hybridoma cell lines grew well after continuous culture for more than three months or after being stored in liquid nitrogen for six months, and the titers of secreting McAbs were stable. Conclusion： The McAbs against respiratory syncytial virus are obtained. It lay a foundation for early diagnosis and further study of RSV.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2007年第11期1015-1018,共4页
Chinese Journal of Immunology
关键词
呼吸道合胞病毒
单克隆抗体
杂炎瘤细胞株
Respiratory syncytial virus （RSV）
Monoclonal antibody
Hybridoma cell lines
