H9N2亚型禽流感病毒单克隆抗体的制备及其表位鉴定

Preparation of H9N2 Subtype Avian Influenza Virus Antibodies and Epitope Identification

[目的]制备H9N2亚型禽流感病毒单克隆抗体和鉴定表位。[方法]选取H9N2亚型禽流感病毒(AIV)WD-1株的纯化抗原免疫BALB/c小鼠,对获得2株抗H9N2亚型禽流感病毒的特异性单抗4C10和6E3表位进行鉴定。[结果]单抗4C10和6E3分别属于IgG1和IgG2b亚型,ELISA检测效价分别为1∶10~3和1∶10~5;HI效价分别为2^(15)和2^(14);2株单抗均具有鸡胚中和活性。利用噬菌体展示表位技术对2株单抗的抗原表位进行鉴定,结果显示均为针对HA蛋白的1个线性表位。[结论]该研究为禽流感病毒快速诊断方法的建立提供了依据。

[ Objective] To prepare H9N2 subtype avian influenza virus antibodies and identify epitope. [ Method] The epitopes of two monoclonal antibodies （McAbs） specific to H9 subtype avian influenza virus （AIV） were identified by cell fusion with the BALB/c mice immunized with pttrified H9N2 AIV WD-1. [ Result]The 4C10 and 6E3 were belonged to IgG 1 and IgG 2b,repectively. The ELISA titers were 1：10^3 and 1：10^5. The HI titers in their ascites were 2^15 and 2^14. Two monoclonal antibodies had chicken embryo neutralizing activity. The antigen epitopes of two monoclonal antibodies were all linera epitopes targeting HA protein. [ Conclusion ] The study laid the foundation for the establishment of rapid diagnostic method of avian influenza virus.