1. Characterization of a Novel Anti-Human HB-EGF Monoclonal Antibody Applicable for Paraffin-Embedded Tissues and Diagnosis of HB-EGF-Related Cancers
- Author
-
Yoshiro Kishi, Jun-ichi Akatsuka, Ken-ichiro Ono, Eisuke Mekada, Mika Takagi, and Ryo Iwamoto
- Subjects
0301 basic medicine ,medicine.drug_class ,Heparin-binding EGF-like growth factor ,Immunology ,Biology ,Monoclonal antibody ,Immunofluorescence ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Epidermal growth factor ,Cell Line, Tumor ,Neoplasms ,medicine ,Immunology and Allergy ,Animals ,Humans ,medicine.diagnostic_test ,Epidermal Growth Factor ,Antibodies, Monoclonal ,Original Articles ,Molecular biology ,Immunohistochemistry ,ErbB Receptors ,030104 developmental biology ,Epitope mapping ,Paraffin ,030220 oncology & carcinogenesis ,biology.protein ,Antibody ,Clone (B-cell biology) ,hormones, hormone substitutes, and hormone antagonists ,Epitope Mapping ,Heparin-binding EGF-like Growth Factor - 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.
- Published
- 2016