Mapping of the binding sites of human diamine oxidase (DAO) monoclonal antibodies.

ABSTRACT:

Objective

Recently we characterized five mouse monoclonal antibodies that allow the specific and sensitive detection of human diamine oxidase (DAO). To understand differences in binding characteristics and recognition of enzyme variants, we mapped the antibody binding sites.

Methods

Fragments of human DAO were expressed as glutathione-S-transferase fusion proteins that were used for testing antibody binding on immunoblots. Combined information from species cross-reactivity, sequence comparison and binding site-prediction software were used to localize the epitope recognized by each antibody.

Results

All five monoclonal DAO antibodies bound to linear epitopes between the N3 and enzymatic domains of the 732 amino acid protein. The binding sites could be mapped onto amin

SUBMITTER: Schwelberger HG 

PROVIDER: S-EPMC5807474 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature