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Characterazation of the anti-HEV ORF2 monoclonal antibodies by biosensor

Aim To prepare the anti HEV ORF2 monoclonal antibodies (mAbs), and idantify their partial properties by biosensor. Methods The BALB/c mice were immunized with purified recombinant protein NE2 of HEV ORF2. Anti HEV ORF2 mAbs were prepared by hybridoma technique, and the relative properties of the mAbs were identified by ELISA, Western blot and biosensor. Results 12 hybridoma cell lines steadily secreting the anti NE2 mAbs were obtained. Of these mAbs, mAb 3F5 was IgM, and the others were IgG (IgG1, IgG2a or IgG2b), and the isotype of their light chains were kappa (κ). MAb 3F5 cross reacted to HRP GAM IgG2a and HRP GAM IgM in ELISA, but this mAb was proved to be IgM by biosensor. The results of the Western blot showed that 12 mAbs reacted to NE2 protein. Reactivities of mAbs 1F6, 8C10, 10D4, 12G8, 2C9, 8C11, 8H3 and 13D8 to NE2 dimer and polymer were stronger than that to NE2 monomer, while mAbs 3F5, 7E8, 15B2, and 16D7 had the same reactivities to monomer, dimer or polymer of NE2. The K D of four mAbs (8C11,8H3,13D8 and 16D7) were calculated through binding curve of coulpled NE2 to varying concentritions of mAbs. And from the curve of the binding and dissociation, we found that the binding of mAb 8H3 to NE2 and dissociation from it were much faster than that of the other three. Conclusion Twelve hybodoma cell lines secreting mAbs to recombinant HEV ORF2 protein were obtained and characterized.