Preparation and characterization of monoclonal antibodies against botulinum neurotoxins serotype A / 军事医学

ABSTRACT

Objective To prepare and characterize specific monoclonal antibodies( McAbs) against the heavy chain of botulinum neurotoxin serotype A ( BoNT/AHc ) .Methods BALB/c mice were immunized with purified BoNT/AHc protein.After the fusion of mouse splenic cells with SP2/0 cells, hybridoma cell lines secreted McAbs against BoNT/AHc. The McAbs obtained were characterized by indirect ELISA, Western blotting and rapid isotypingassay before being used in ELISA to detect interaction sites in McAbs and BoNT/AHc preliminarily.Results Antigen protein BoNT/AHc of high purification was obtained.Four hybridoma cell lines secreting McAbs against BoNT/AHc were screened,named 1A4,3H3, 3H7 and 5H8,respectively.Their titers of McAbs were all above 3.0 ×103 .They were specifically combined with BoNT/AHc protein by Western blotting.The isotype of 1A4 and 3H7 was IgG1(Κ),that of 3H3 was IgM(Κ),and that of 5H8 was IgG2b(Κ).Additive ELISA showed that epitopes recognized by the four McAbs were close.ELISA analysis confirmed the interaction epitopes in McAbs and BoNT/AHc.Conclusion Monoclonal antibodies against BoNT/AHc are prepared and characterized,providing effective tools for studying the neutralizing antibody and antibody epitopes of BoNT/AHc.