title: Characterization of a Novel Anti-Human HB-EGF Monoclonal Antibody Applicable for Paraffin-Embedded Tissues and Diagnosis of HB-EGF-Related Cancers.
abstract: Heparin-binding EGF-like growth factor (HB-EGF) is a member of the EGF family of growth factors that bind to and activate the EGF receptor (EGFR/ErbB1) and ErbB4. HB-EGF plays pivotal roles in pathophysiological processes, including cancer. Thus, monoclonal antibodies (mAbs) for HB-EGF detection could be an important tool in the therapeutic diagnosis of HB-EGF-related cancers and other diseases. However, few mAbs, especially those applicable for immunohistochemistry (IHC), have been established to date. In this study, we generated a clone of hybridoma-derived mAb 2-108 by immunizing mice with recombinant human HB-EGF protein expressed by human cells. The mAb 2-108 specifically bound to human HB-EGF but not to mouse HB-EGF and was successful in immunoblotting, even under reducing conditions, immunoprecipitation, and immunofluorescence for unfixed as well as paraformaldehyde-fixed cells. Notably, this mAb was effective in IHC of paraffin-embedded tumor specimens. Epitope mapping analysis showed that mAb 2-108 recognized the N-terminal prodomain in HB-EGF. These results indicate that this new anti-HB-EGF mAb 2-108 would be useful in the diagnosis of HB-EGF-related cancers and would be a strong tool in both basic and clinical research on HB-EGF.
authors.affiliation: 1 Department of Cell Biology, Research Institute for Microbial Diseases, Osaka University , Osaka, Japan .
authors.forename: Ryo
authors.initials: R
authors.lastname: Iwamoto
authors.affiliation: 1 Department of Cell Biology, Research Institute for Microbial Diseases, Osaka University , Osaka, Japan .
authors.forename: Mika
authors.initials: M
authors.lastname: Takagi
authors.affiliation: 2 Medical & Biological Laboratories Co., Ltd , Nagoya, Japan .
authors.forename: Jun-Ichi
authors.initials: J
authors.lastname: Akatsuka
authors.affiliation: 2 Medical & Biological Laboratories Co., Ltd , Nagoya, Japan .
authors.forename: Ken-Ichiro
authors.initials: K
authors.lastname: Ono
authors.affiliation: 2 Medical & Biological Laboratories Co., Ltd , Nagoya, Japan .
authors.forename: Yoshiro
authors.initials: Y
authors.lastname: Kishi
authors.affiliation: 1 Department of Cell Biology, Research Institute for Microbial Diseases, Osaka University , Osaka, Japan .
authors.forename: Eisuke
authors.initials: E
authors.lastname: Mekada
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D045744:Cell Line, Tumor; D004815:Epidermal Growth Factor / Q000276:immunology* / Q000302:isolation & purification; D018604:Epitope Mapping; D066246:ErbB Receptors / Q000276:immunology / Q000302:isolation & purification; D066257:Heparin-binding EGF-like Growth Factor / Q000276:immunology* / Q000302:isolation & purification; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D009369:Neoplasms / Q000175:diagnosis / Q000276:immunology*; D010232:Paraffin
chemical_list: D000911:Antibodies, Monoclonal; C587883:HBEGF protein, human; D066257:Heparin-binding EGF-like Growth Factor; D004815:Epidermal Growth Factor; D010232:Paraffin; D066246:ErbB Receptors