Production And Characterization Of The Monoclonal Antibody Against Capsid Protein Of Avian Hepatitis E Virus

Hepatitis E (HE) is a kind of acute viral hepatitis caused by avian Hepatitis E virus (HEV), which is transmited by intestinal tract. Avian HEV can cause hepatitis-splenomegaly syndrome, hepatonecrosis, hepatic hemorrhage, regressive ovaries and upgrade mortality.In 2001, Haqshenas isolated avian hepatitis E virus (avian HEV) from biles of chickens that had hepatitis-splenomegaly (HS) syndrome. There have been several reports about avian HEV serological invetigation after avian HEV had been discovered. But, avian HEV serological investigation is located in in primary level because of cross-reactivity of antibodies against ORF2 of avian, swine and human HEVs. The ORF2 gen of avian HEV shares approximately 48-49% amino acid sequence with those of human and swine HEVs. The most conserved region of the ORF2 protein is located at aa339-606.In this study, we produced six monoclonal antibodies (Mabs) against avian HEV ORF2-1 protein, designated 7H12, 3E8, 1B5, 3A1 and 6G8, after standard operation procedures of expression of ORF2-1, immune mouse, cell fusion and subclone. Six monoclonal antibodies (Mabs), which recognize six epitopes aa339-355, aa355-384, aa355-384, aa384-414, aa389-398 and aa510-516, were screened and established according to ELISA results between 9 truncated ORF2 proteins, 8 peptides and Mabs. Mabs 3E8 and 1B5 recognize different epitopes which were located at aa355-384 according to competitive ELISA between HRP-3E8 and 1B5.In 2009, Zhao Qin isolated avian hepatitis E virus (avian HEV) from biles of chickens that had hepatitis-splenomegaly (HS) syndrome in China, injected Chinese avian HEV (CaHEV) to SPF chickens and collected fecal, serum samples. Mabs were used for establishing immune capture of Chinese avian HEV (CaHEV), Mabs 7H12 and 1H5 of six Mabs can capture avian HEV particeles, which were coincidence with the results of immunohistochemical.In this study, six Mabs provide material basis for search of serological investigation among avian avian, swine and human HEVs. Moreover, Mabs 7H12 and 1H5 provide theoretical basis for avian HEV diagnosis.