Interaction of the octapeptide angiotensin II with a high-affinity single-chain Fv and with peptides derived from the antibody paratope
| Autor: | Thérèse Combes, Daniel Laune, Dominique Simon, Jean Claude Mani, Gabriel Badouaille, Martine Pugnière, Isabelle Teulon, Claude Granier, Pascale Cohen |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
medicine.drug_class
Molecular Sequence Data Immunology Antibody Affinity Immunoglobulin Variable Region Gene Expression Immunoglobulin light chain Monoclonal antibody Antigen Antibody Specificity Escherichia coli medicine Immunology and Allergy Single-chain variable fragment Amino Acid Sequence Immunoglobulin Fragments Base Sequence biology Chemistry Angiotensin II Molecular biology Recombinant Proteins Solubility Biochemistry Biotinylation biology.protein Immunoglobulin Light Chains Paratope Antibody Immunoglobulin Heavy Chains Oligopeptides Sequence Alignment |
| Zdroj: | Journal of Immunological Methods. 254:147-160 |
| ISSN: | 0022-1759 |
| Popis: | The amino-acid sequence of the very high-affinity anti-angiotensin II monoclonal antibody 4D8 was predicted from the nucleotide sequence of the heavy and light chain variable genes. The single-chain variable fragment (scFv) was constructed and expressed in Escherichia coli as a soluble protein and at the surface of the filamentous M13 phage and was compared with the full-length antibody (Ab). The scFv showed the same specificity profile and affinity constant as the intact antibody (5.0×10 10 and 8.0×10 10 M −1 , respectively, by Scatchard analysis). Several peptides from the set of overlapping dodecapeptides covering the variable domains of 4D8 mAb were found to specifically bind biotinylated angiotensin II: peptides from the L1, L2, L3 and H1 regions had the strongest capacity to bind the antigen. |
| Databáze: | OpenAIRE |
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