| In this experiment, several hybridoma cell strains were developed by fusion of SP2/0 mouse myeloma cells with spleen cells isolated from a BALB/C mouse which was immunized with the purified AIV DZJ/1100.Three McAb cell strains were determined by hemagglutinin inhibition (HI): DD7, FG10, and CB12. Two McAb cell strains were determined by indirect ELISA coated with purified virus: EC6, EA3. Two McAb cell strains were determined by indirect ELISA coated with recombinant NP protein of AIV: 4H1, 4H7. The titers of ascites: the specificial McAbs of HA (HI titers) were: 213, 215, 211, the specificial McAbs of NP(ELISA titers) were: 1:16000, 1:32000. the specificial McAbs of other proteins on AIV (ELISA titers) were: 1:32000, 1:64000.The subtypes of HA McAbs belong IgG1, IgG2a and IgG2a.; The subtypes of Np McAbs belong IgM and IgM, The subtypes of other proteins McAbs on AIV belong IgG1 and Ig2a. all McAbs are the k strains. All McAbs had good stability of acid,basic and heat.The results of HI cross-reaction and neutralization test suggest that the McAbs FG10 and CB12 had excellent reactivity with others H5 subtype viruses, but DD7 had no cross-reactivity and neutralization to DKZJ/0202. These data imply there is no epitope site which recognized by DD7 on the HA of DKZJ/0202. It is interesting that DD7 had high HI titer to DKSH/3502, but fail to neutralize the DKSH/3502virus in chicken egg embryo. Some may be concluded that the HA epitopes is not same as the sites of neutralization in AIVs. The specificity result of H5 McAbs which react to others HA subtype (H1-H4, H6-H15) AIV by HI method suggest the three McAbs had good specificity to H5 AIV and can be used to verdict the H5 subtype AIVs from suspicious HPAI case.Candidate: Zhang LichunSpeciality: Preventive Veterinary ScienceSupervisor: Associate Prof. Jia Yongqing...
|
PDF Full Text Request