摘要
目的:获得能稳定分泌抗人呼吸道合胞病毒(human respiratory syncytial virus,RSV)融合糖蛋白(fusion glycoprotein,F)单克隆抗体(monoclonal antibody,mAb)的杂交瘤细胞株,以期用于RSV感染的早期诊断和被动免疫治疗研究。方法:通过杂交瘤技术制备可特异性识别RSV F的单抗,体外鉴定生物学特性。结果:获得了可分泌抗RSV F蛋白的杂交瘤细胞株F8,体外连续传代培养2个月,能稳定分泌抗体F8,培养上清效价为1∶1000,亲和常数(Ka)为6.8×108L/mol。F8属IgG1型抗体,可特异性识别RSV F1亚单位的AA 205-222。免疫酶法蚀斑减少中和实验证实F8具有体外中和活性及融合抑制活性。结论:获得具有中和活性的抗RSV F蛋白的单克隆抗体,为RSV感染的早期诊断及被动免疫治疗等奠定了基础。
Objective: To obtain the hybridoma cell line stably secreting monoclonal antibody (mAb) specific for F glycoprotein (F) of human respiratory syncytial virus (RSV) for the further study of early detection and passive immunotherapy against RSV infection. Methods: The mAbs against F were prepared by hybridoma technology and then the associated characteristics were identified in vitro. Results : The hybridoma cell line of F8 capable of stably secreting mAb was established. It grew well after continuous culture for two months, the resultant supernatant titer of the mAb is 1 : 1000. The mAb is IgG1 and its relative affinity (Ka) is 6.8 ~ 108 L/tool. The potential epitope recognized by F8 is located around AA 205-222 of F1 subunit of RSV F. Through the immunoenzyme method, the neutralizing activity and fusion inhibition capacity of mAb F8 in vitro were also confirmed. Conclusion: The mAb F8 against RSV F with neutralizing activity was obtained. It lays a foundation for early diagnosis and passive immunotherapy of RSV infection.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2012年第2期63-68,共6页
China Biotechnology
基金
北京市自然科学基金资助项目(7092053)
关键词
人呼吸道合胞病毒
融合糖蛋白
单克隆抗体
杂交瘤技术
Human respiratory syncytial virus Fusion glycoprotein Monoclonal antibody Hybridomatechnology