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Anti-human HB-EGF monoclonal antibodies inhibiting ectodomain shedding of HB-EGF and diphtheria toxin binding.
- Source :
-
Journal of biochemistry [J Biochem] 2010 Jul; Vol. 148 (1), pp. 55-69. Date of Electronic Publication: 2010 Mar 23. - Publication Year :
- 2010
-
Abstract
- HB-EGF is a member of the EGF family of growth factors that bind and activate the EGF receptor. HB-EGF is synthesized as a membrane-anchored protein (proHB-EGF), and then proteolytically cleaved, resulting in the mitogenically active soluble form. ProHB-EGF functions as the receptor for the diphtheria toxin (DT). HB-EGF plays pivotal roles in pathophysiological processes, including cancer. Monoclonal antibodies (mAbs) specific for HB-EGF could be an important tool in HB-EGF research. However, few such mAbs have been established to date. In this study, we newly generated seven clones of hybridoma-derived mAbs by immunizing HB-EGF null mice with recombinant human HB-EGF protein. All mAbs specifically bound to human HB-EGF but not to mouse HB-EGF. Epitope mapping analysis showed that most of the mAbs recognized the EGF-like domain. Although none of the newly isolated mAbs directly inhibited the mitogenic activity of HB-EGF for EGFR-expressing cells, some strongly inhibited DT-binding. Interestingly, some of the mAbs efficiently inhibited ectodomain shedding of proHB-EGF, and consequently prevented the cell growth of the EGFR-expressing cells in a co-culture system with proHB-EGF-expressing cells. Hence, these new anti-HB-EGF mAbs may advance clinical as well as basic research on HB-EGF.
- Subjects :
- Amino Acid Sequence
Animals
Antibodies, Monoclonal immunology
Antibody Specificity
Cell Line
Cell Membrane metabolism
Cell Proliferation
Coculture Techniques
Epitope Mapping
Heparin-binding EGF-like Growth Factor
Humans
Immunization
Immunoassay
Immunoprecipitation
Mice
Mitogens metabolism
Molecular Sequence Data
Neutralization Tests
Protein Binding
Protein Structure, Tertiary
Antibodies, Monoclonal biosynthesis
Diphtheria Toxin metabolism
Intercellular Signaling Peptides and Proteins chemistry
Intercellular Signaling Peptides and Proteins immunology
Subjects
Details
- Language :
- English
- ISSN :
- 1756-2651
- Volume :
- 148
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Journal of biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 20332144
- Full Text :
- https://doi.org/10.1093/jb/mvq033