IDENTIFICATION OF MAJOR ANTIGENIC DOMAINS OF DUCK HEPATITIS-B VIRUS PRE-S PROTEIN BY PEPTIDE SCANNING

Neutralization epitopes of duck hepatitis B virus (DHBV) have been previously mapped within the N-terminal portion of the pre-S protein using monoclonal antibodies. However, the immune response of ducks to this region is not well characterized at the amino acid level. To this end, we have immunized adult Pekin ducks with either DHBV positive serum or bacterially expressed DHBpre-S polypeptide representing the N-terminal portion of the DHBV pre-S region. We have demonstrated that adult ducks inoculated with either antigen developed antibodies to the DHBV pre-S region starting 5 to 10 days postinjection. The sera of all ducks, irrespective of the immunogen used, exhibited a significant protective activity against DHBV, as assessed in vivo. To identify which pre-S domains bind antibodies from these duck sera, we have used the Pepscan methodology with overlapping octapeptides spanning the DHBV pre-S sequence from amino acids 1 to 145. Using this approach, five major antigenic domains, (7)KSMDVRRI(14), (22)NQLAGRMIP(30), (58)TLQNQGAW(65), (71)RRVGLSNPT(74), and (127)GDDPLLGNQ(135) were identified within the DHBV pre-S region. (C) 1994 Academic Press, Inc.