小鼠抗人呼吸道合胞病毒磷蛋白(P蛋白)单克隆抗体的制备和鉴定

Preparation and identification of mouse anti-human monoclonal antibody against P protein of respiratory syncytial virus

目的制备抗人类呼吸道合胞病毒(HRSV)磷蛋白(P蛋白)单克隆抗体,并对其进行鉴定。方法以原核表达系统表达的HRSV P蛋白重叠肽段作为免疫原,采用杂交瘤技术筛选P蛋白单克隆抗体,Western blot法验证筛选获得的单克隆抗体与P蛋白的结合活性,免疫荧光细胞化学染色确定获得的单克隆抗体能否用于RSV感染后HEp-2细胞中P蛋白的检测。结果筛选出反应性好、特异性识别HRSV P蛋白的单克隆抗体P181-15A3、P211-16D8。获得的单克隆抗体通过Western blot法验证可与P蛋白结合,并可用于RSV感染后HEp-2细胞中P蛋白的免疫细胞化学检测。结论成功制备了小鼠抗HRSV P蛋白的单克隆抗体。

Objective To prepare the monoclonal antibodies specifically against P protein of human respiratory syncytial virus(HRSV)and identify it.Methods HRSV P protein prepared by prokaryotic expression in the form of overlapping peptides was used as the immunogen,and monoclonal antibodies(mAbs)were screened by hybridoma technology.Western blot analysis was used to verify the binding activity of the screened mAbs and P protein,and immunofluorescence cytochemical staining was used to determine whether the obtained mAbs could be used to detect the expression of P protein in HEp-2 cells infected with HRSV.Results P181-15A3 and P211-16D8 with great reactivity and specific recognition of HRSV P protein were screened.Both mAbs could bind to P protein by Western blot analysis and could be used for immunocytochemical detection of P protein in HEp-2 cells after HRSV infection.Conclusion We have successfully prepared the mAbs against HRSV P protein.