title: Multiplex peptide-based B cell epitope mapping.
abstract: B cell epitope mapping is widely applied to determine antibody-binding sites. Several methods exist to map B cell epitopes and here we describe three methods that are characterized by the simultaneous analysis of multiple peptides. In the first approach a microarray of overlapping synthetic peptides derived from an antigenic protein is used and the binding of the antibodies is analyzed by fluorescently labeled secondary antibodies. This method is particularly suited for the identification of linear epitopes of an established target protein. In the second approach the binding of antibodies to a random synthetic peptide library immobilized on microbeads is determined by enzyme-conjugated secondary antibodies and the selection of antibody-bound beads by a light microscope. This method can be applied when information on the identity of the antigenic protein is lacking. In the third method an antigen is proteolytically digested and antibody binding to the resulting peptides is analyzed by surface plasmon resonance imaging (iSPR). The latter method can be applied when the purified antigenic protein is available.
authors.affiliation: Department of Biomolecular Chemistry, Radboud Institute for Molecular Life Sciences, Institute for Molecules and Materials and Netherlands Proteomics Centre, Radboud University Nijmegen, 9101, NL-6500 HB, Nijmegen, The Netherlands.
authors.forename: Sanne M M
authors.initials: SM
authors.lastname: Hensen
authors.forename: Merel
authors.initials: M
authors.lastname: Derksen
authors.forename: Ger J M
authors.initials: GJ
authors.lastname: Pruijn
mesh_terms: D018604:Epitope Mapping / Q000379:methods*; D018985:Epitopes, B-Lymphocyte / Q000032:analysis / Q000276:immunology*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000032:analysis / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology / Q000302:isolation & purification; D019151:Peptide Library; D010449:Peptide Mapping / Q000379:methods; D040081:Protein Array Analysis / Q000379:methods
chemical_list: D018985:Epitopes, B-Lymphocyte; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D019151:Peptide Library