title: TD-12 workshop report: characterization of monoclonal antibodies to neuron-specific enolase.
abstract: Twelve antibodies to neuron-specific enolase (NSE) have been evaluated by four working groups. Human brain γγ-enolase, neuroblastoma-derived αγ-enolase, and recombinant γγ-enolase were used to determine antibody specificity and binding kinetics. All antibodies were found to be specific for the γ-isoform. It was possible to assign 11 of the antibodies to at least five epitope groups based on cross-inhibition experiments, QCM and SPR technology, and immunoassay combinations. Antibodies 9601 and 9602 showed the highest affinity for both native and recombinant γγ-enolase. Immunometric assays for both γγ- and αγ-enolase could be made by pairing 9601 with most of the other ISOBM antibodies. Antibodies differed in their ability to recognize native αγ-enolase, native γγ-enolase, and recombinant γγ-enolase. Some immunometric assay combinations appear to favor the detection of heterodimeric αγ-enolase over the homodimeric γγ-enolase. Although the majority of the antibodies failed to detect human NSE or recombinant NSE in Western blots, mAb 9601 recognized both, while E17 and 18E5 were specific for human NSE.
authors.affiliation: Department of Medical Biochemistry, Norwegian Radium Hospital, Oslo University Hospital, Montebello, 0310, Oslo, Norway. elisabeth.paus@klinmed.uio.no
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Paus
authors.forename: Klaus
authors.initials: K
authors.lastname: Hirzel
authors.forename: Maria
authors.initials: M
authors.lastname: Lidqvist
authors.forename: Matti
authors.initials: M
authors.lastname: Höyhtyä
authors.forename: David J
authors.initials: DJ
authors.lastname: Warren
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D015153:Blotting, Western; D006801:Humans; D007527:Isoenzymes; D010751:Phosphopyruvate Hydratase / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D010751:Phosphopyruvate Hydratase