The Antigen-antibody Immune Preparation Of Hepatitis B E Antigen-specific Epitope Antibody And Detection Applications

Due to its apparent practical value, hepatitis B virus e antigen(HBeAg) hasattracted increasing attention. Detection of HBeAg in the serum of individualsinfected with hepatitis B virus (HBV) can indicates both a high infectivity of theserum and a poor prognosis of HBV-related diseases in the host. Variousimmunoassays for the detection of HBeAg were developed, although development offormat and components continues in order to increase the sensitivity and specificity ofthe assays.In this study, we report an antigen-antibody co-immunization way of raising theanti-HBe antibodies as well as some characterizations and application work of theantibodies. We immunize with both recombinant HBeAg and its predominantantibody named ewb, which recognized the dominant antigenic determination ofHBeAg(HBe βepitope, 128-133 amino acids). This new way of immunization isimportant and necessary because using the traditional way we failed to raise anantibody against the different epitope of HBeAg. Mostly, we get the antibodiessimilar to ewb, then it is difficult to construct pair-Abs to be used in serologicaldiagnosis. The results suggested that the co-immunization strategy is feasible to raisean antibody(S-73-2) which is different from the predominant antibody——ewb andcan be used as pair-Abs. We further map its epitope with both truncated HBeAgfragments and synthesis peptides, founding that the S-73-2 is a conformationalantibody recognizing the N-terminal of HBeAg. Using the S-73-2 in a sandwichELISA-kit, we specifically detect the recombinant HBeAg suggesting that this kit hada potential application in clinical use.