Preparation and identification of antibodies against respiratory syncytial virus nucleoprotein
10.3760/cma.j.issn.0254-5101.2018.03.001
- VernacularTitle:呼吸道合胞病毒核蛋白检测抗体的筛选与鉴定
- Author:
Xue LIN
1
,
2
;
Wei ZHANG
;
Min ZHAO
;
Zizheng ZHENG
;
Ningshao XIA
Author Information
1. 361102 厦门大学,分子疫苗学与分子诊断学国家重点实验室,国家传染病诊断试剂与疫苗工程技术研究中心
2. 361102 厦门大学,公共卫生学院
- Keywords:
Respiratory syncytial virus;
Nucleoprotein;
Monoclonal antibodies
- From:
Chinese Journal of Microbiology and Immunology
2018;38(3):161-168
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare and identify the monoclonal antibodies against respiratory syn-cytial virus nucleoprotein(RSV N protein). Methods A prokaryotic expression system was used to express recombinant RSV N protein in Escherichia coli (E.coli). BALB/c mice were immunized with the recombi-nant N protein after purification. Monoclonal antibodies (McAbs) against the N protein were sorted from these BALB/c mice and then were further characterized by Western blot, ELISA and immunofluorescence. Furthermore,this study used orthogonal experiment to identify McAbs pairs,which could be used for diagno-sis. Results This study succeeded in obtaining 24 hybridoma cells that stably secreted monoclonal antibod-ies against RSV N protein. These antibodies showed good reactivity in ELISA,of which eight had strong spe-cificity in Western blot and 13 could be used in immunofluorescence. This study obtained two McAbs pairs (12F2/11H8-HRP and 12F2/15A8-HRP) that could be used in RSV detection. Conclusion This study succeeded in screening and preparation of McAbs against RSV N protein and obtaining two potential McAbs pairs for rapid detection of RSV.
