Abstract: Objective To predict and identify B-cell linear epitopes of hepatitis B e antigen (HBeAg). Methods The B-cell linear
epitopes of HBeAg were predicted using the software provided by NCBI Database and Immune Epitope Database (IEDB) and
synthesized by a solid-phase method followed by conjugation with keyhole limpet hemocyanin (KLH). The KLH conjugates
were used for immunization of New Zealand white rabbits, and the immune response of the rabbits was monitored by direct
ELISA using a bovine serum albumin conjugate of the predicted epitopes. Results Four new B-cell linear epitopes of HBeAg
were identified, namely 1MDIDPYKEFG10, 37LYREALESPEHCSP50, 74SNLEDPAS81 and 127RTPPAYRPPNAPIL140. The rabbits
immunized with the KLH conjugate showed an antibody titer over 1∶512 000. The antisera of B-cell linear epitopes collected
could specifically react with HBeAg as shown by ELISA. Conclusion Four B-cell linear epitopes of HBeAg have been
confirmed using bioinformatics methods, which provides new evidence for further functional studies of HBeAg in hepatitis B.