Application of the Spot method to the identification of peptides and amino acids from the antibody paratope that contribute to antigen binding
| Autor: | Francois Rieunier, Thierry Chardès, Sylvie Villard, Cédric Bès, Franck Molina, Daniel Laune, Gaëlle Ferrières, Claude Granier |
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| Rok vydání: | 2002 |
| Předmět: |
Models
Molecular Antigen-Antibody Complex Protein Conformation medicine.drug_class Amino Acid Motifs Molecular Sequence Data Immunology Peptide Monoclonal antibody Antigen-Antibody Reactions Mice medicine Animals Humans Immunology and Allergy Amino Acid Sequence Antigens chemistry.chemical_classification Binding Sites Hybridomas Sequence Homology Amino Acid biology Antibodies Monoclonal Biological activity Amino acid chemistry Biochemistry biology.protein Paratope Identification (biology) Binding Sites Antibody Antibody |
| Zdroj: | Journal of Immunological Methods. 267:53-70 |
| ISSN: | 0022-1759 |
| DOI: | 10.1016/s0022-1759(02)00140-0 |
| Popis: | Overlapping peptide scans prepared by Spot synthesis have been used to map interaction sites in several systems. Here we report our experience with this approach to identify peptides from the variable parts of anti-hapten, anti-peptide and anti-protein antibodies that retain their specific antigen-binding capacity in the Spot format. In general, the identification by the Spot method of antigen-reactive peptides was confirmed by using soluble peptides which demonstrated antigen-binding capacity in ELISA or Biacore and, biological activity for some peptides derived from anti-CD4 antibodies. The Spot method was also used to map precisely key residues from the antibody paratope. The identification of critical residues from an anti-troponin I antibody of diagnostic interest is reported as well as the compiled results from the analysis of five other antibodies of various specificities. A critical assessment of our results is provided by comparing results obtained by our approach in the mapping of antibody residues critical for antigen binding with data from the literature concerning the structural analysis of antigen-antibody complexes. |
| Databáze: | OpenAIRE |
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