Prokaryotic expression and identification of B- and T-cell combined epitopes of Em95 antigen of Echinococcus multilocularis
Related Experiment Video
Updated: Apr 24, 2026
Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope
Published on: March 24, 2017
Hongying Wang1, Fengbo Zhang1, Xiumin Ma1
1State Key Laboratory Incubation Base of Xinjiang Major Diseases Research, The First Affiliated Hospital of Xinjiang Medical University Urumqi 830054, Xinjiang, China.
Objective:
This study is to clone and identify B- and T-cell combined epitopes from Em95 antigen.
Methods:
The B- and T-cell combined epitopes were predicted using bioinformatic software. Two DNA sequences of Em95-1 (which contained the coding region of one B- and T-cell combined epitope) and Em95-2 (which contained the coding regions of two B- and T-cell combined epitopes) were amplified by PCR. Em95-1 and Em95-2 were cloned into pET32a vector for protein expression. Rabbit was immunized with the expressed proteins of rEm95-1 and rEm95-2 to produce polyclonal antibodies. The immunogenicity and antigenicity of rEm95-1 and rEm95-2 were examined by Western blot analysis.
Results:
The three B- and T-cell combined epitopes were successfully cloned and expressed in PET32a vector. The recombinant antigens of rEm95-1 and rEm95-2 could specifically bind the human serum from patients with alveolar echinococcosis and specifically bind the prepared polyclonal antibodies.
Conclusion:
Three B- and T-cell combined epitopes were successfully cloned with good immunogenicity and antigenicity. Our data suggest that B- and T-cell combined epitopes predicted from the Em95 antigen may be used for the construction of high-valence vaccines and as targets for prevention of echinococcosis.
