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A model system for detection and isolation of a tumor cell surface antigen using antibody phage display
- Source :
- Journal of Immunological Methods. 203:11-24
- Publication Year :
- 1997
- Publisher :
- Elsevier BV, 1997.
-
Abstract
- To establish a screening procedure for tumor cell-surface reactive Fabs, we used a model antigen/antibody system including the epidermal growth factor receptor (EGF-R) and the anti-EGF-R monoclonal antibody 425. The 425 Fab was displayed on the surface of M13 filamentous phage. In a screening assay for 425 phage binding to tumor cell surfaces, biotinylated 425-phage bound specifically to EGF-R-positive A431 epidermoid carcinoma cells and not to K562 non-expressor erythroleukemia cells. With a model library, the sensitivity of phage enrichment by phage binding to cell surfaces was one 425-phage in 20,000 unrelated phages after 4 rounds of panning on A431 cells. In a phage tissue screening assay, 425-phage, but not unrelated phage, bound specifically to melanoma cells expressing EGF-R. Epitope and idiotope specificity of 425-phage was demonstrated in phage competition assays, using as targets A431 cells and anti-idiotypic antibodies to monoclonal antibody 425, respectively. Finally, the EGF-R protein was directly isolated from A431 cell extracts, using biotinylated 425-phage. The data obtained with the 425 model library system demonstrate the usefulness of antibody phage display for the rapid identification and isolation of tumor or other disease-related cell surface antigens.
- Subjects :
- Phage display
viruses
Immunology
Adenocarcinoma
Epitope
Immunoglobulin Fab Fragments
Antigen
Antigens, Neoplasm
Tumor Cells, Cultured
Humans
Immunology and Allergy
Melanoma
biology
Models, Immunological
Antibodies, Monoclonal
Idiotopes
Molecular biology
Antibodies, Anti-Idiotypic
ErbB Receptors
Epidermoid carcinoma
Biotinylation
Antigens, Surface
biology.protein
Binding Sites, Antibody
Antibody
A431 cells
hormones, hormone substitutes, and hormone antagonists
Bacteriophage M13
Protein Binding
Subjects
Details
- ISSN :
- 00221759
- Volume :
- 203
- Database :
- OpenAIRE
- Journal :
- Journal of Immunological Methods
- Accession number :
- edsair.doi.dedup.....9886d2f42b48e1057e13d371c0d5eae6
- Full Text :
- https://doi.org/10.1016/s0022-1759(97)00005-7