抗呼吸道合胞病毒单克隆抗体的制备及其鉴定

Preparation and identification of monoclonal antibody against respiratory syncytial virus

目的制备抗呼吸道合胞病毒（ＲＳＶ）的单克隆抗体，并对其特性进行系统地鉴定。方法以活的ＲＳＶＬｏｎｇ株滴鼻（ＴＣＩＤ５０为１×１０－６，取１００ＴＣＩＤ５００１２５ｍｌ进行）加ＲＳＶ感染Ｈｅｐ２细胞（ＣＰＥ达～）腹腔注射免疫Ｂａｌｂ／ｃ鼠，用细胞融合技术制备能稳定分泌抗ＲＳＶ的单克隆抗体细胞株。结果细胞融合一次成功，融合率为９６％，抗ＲＳＶ抗体阳性孔率为６５％。鉴定１株杂交瘤细胞Ｆ３株，染色体数为９８条，抗体亚类属ＩｇＧ１，对ＲＳＶ抗原特异；在无补体存在的情况下其腹水抗体中和效价为１∶１２８，且具融合抑制活性，腹水融合抑制效价为１∶６４。结论Ｆ３株单克隆抗体为抗ＲＳＶＦ蛋白的ＭｃＡｂ。

Objective To establish the monoclonal antibody (McAb) against respiratory syncytial virus(RSV) and to identify its properties. Methods BALB/c mice were immunized with live RSV Long strain intranasally, and enhanced with Hep 2 cells infected RSV intraperitoneally. SP2/0 cells were fused with the spleen cells harvested from Balb/c mice. Results Cells were fused successfully only by one time, the fusion rate was 96 percent and the positive rate was 65 percent. F 3 strain hybridoma cells could secret McAb against RSV specifically, the McAb type belonged to IgG 1 subtype. It had activities of neutralization and fusing inhibition and its neutralizing and fusing inhibition titres of ascite fluids of mice were 1∶128 and 1∶64 respectively. Conclusion The antibody of F 3 strain is anti RSV F McAb.