抗人呼吸道合胞病毒融合糖蛋白单克隆抗体的制备及体外鉴定

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6.8×10⁸ L/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。

Primary Studies of Monoclonal Antibody Against Fusion Glycoprotein of Human Respiratory Syncytial Virus

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/mol.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.