Identification of functional interaction sites on proteins using bacteriophage-displayed random epitope libraries
| Autor: | H. Pannekoek, A.J. van Zonneveld, M van Meijer, B. M. van den Berg |
|---|---|
| Přispěvatelé: | Other departments |
| Jazyk: | angličtina |
| Rok vydání: | 1995 |
| Předmět: |
medicine.drug_class
Phagemid Genetic Vectors Molecular Sequence Data Computational biology Monoclonal antibody Coliphages Epitope Antigen-Antibody Reactions Bacteriophage Plasminogen Activator Inhibitor 1 Genetics medicine Humans Amino Acid Sequence Panning (camera) Gene DNA Primers Base Sequence Linear epitope biology cDNA library Antibodies Monoclonal General Medicine biology.organism_classification Molecular biology Recombinant Proteins Epitope Mapping Protein Binding |
| Zdroj: | Gene, 167(1-2), 49-52. Elsevier |
| ISSN: | 0378-1119 |
| DOI: | 10.1016/0378-1119(95)00614-1 |
| Popis: | We describe a phage-display-based method to identify epitopes or interaction sites on proteins. DNA encoding the protein of interest is partially degraded with DNase I to generate random fragments of 50–200 bp. These fragments are then cloned into a phagemid vector that has been modified to allow the expression of the random fragments and the construction of a (bacterio)phage-displayed random epitope library. Phages displaying functional epitopes can be selected from these libraries by affinity selection or panning. To test this method we have constructed a random-epitope library for human plasminogen-activator inhibitor 1 and used this library to map the epitope of a monoclonal antibody (mAb) directed against this protein. By alignment of the selected overlapping epitope-containing fragments, we were able to locate the epitope of the mAb on a stretch of 39 amino acids spanning from E128 to V166. The approach may also be applied to more complex systems than single-protein genes, such as viral genomes or complete cDNA libraries. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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