Development And Characterization Of Monoclonal Antibody Against H5N1 Subtype AIV

In this experiment, using three HPAIV H5 subtype A/duck/Guangdong/S1322/201 0(DK/GD/S1322/2010)A/duck/Xinjiang/S3391/2012(DK/XJ/S3391/2012) as immunogen, six week to eight week female BALB/c mice were immunized. Splenic cells from im munized mouse were fused with SP2/0 cells. After tested with the HI and ELISA, we got six hybridoma cell strains, designated A4?D11?F3?G2?B6 and C8. The HI test showed that titers of cell supernatants and ascites were 23,22,23,24,22,22 and 211,211,213,213,28,28. The heavy chain of six McAbs belong IgG1 and IgM. The lighe chain of six McAbs were the k strains. The average chromosome number of six hybridoma cells was between 102 and 108.In the HI and neutralizing activity test, A4 and D11 reacted well with the H5 subtype AIVs of clade 2.3.2.B and clade 2.3.2.C of DK/GD/S1322/2010, but did not reacted with the H5 subtype AIVs of clade 2.3.4 and clade 7; F3 and G2 only reacted well the H5subtype and N8 sutype of DK/XJ/S3391/2012, but did not reacted with AIVS of other clade 2.3.4 and clade 7; B6 and C8 reacted with part of the H5 subtype AIVs of clade 7 of CK/LN/S4092/2011, but did not reacted with the other clade of H5 subtype AIVs. The McAbs could recognize the antigen site of parental virus, and reflect a certain activity in emnryo neutralization tese. Indirect immunofluorescence assay indicated that six McAbs reacted well with parental virus. A4 and D11 have good reaction with PCAGGS-HA through the western-blot.This results suggest that the MAbs would be useful for the surveillance and diagnosis of H5 subtype AIVs.