Peptide Models of Immunological Recognition: Paratope Dissection by Multiple Peptide Synthesis
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Daniel Laune
Abstract
New approaches to obtain information about the residues of the antibody paratope involved in the interaction with antigen would be very useful to help perform rational mutagenesis of antibodies for improving sensitivity or selectivity in immunoassays. We have evaluated the possibility of dissecting the antibody paratope into large sets of short (12 residues) overlapping peptides to determine the contribution of each peptide to antigen binding. Our results show that the systematic analysis of the antigen-binding properties of heavy chain variable segment and light chain variable segment derived peptides of HyHEL-5, a model anti-lysozyme antibody, can be an experimental approach to the selection of paratope-derived peptides with antigen-binding activity. Detailed analysis of the contribution of each residue from each binding peptide permitted the identification of residues contributing to antigen binding. Of the 38 residues we identified, 22 corresponded to residues that had been shown by X-ray crystallography to be at the interface between HyHEL-5 and lysozyme. The peptide analysis we have developed is thus a way to map the subset of residues from the antibody paratope involved in antigen recognition. The same peptide approach was used to map the idiotope that an anti-idiotypic antibody recognized in the paratope of its cognate antibody.
Walter de Gruyter GmbH & Co. KG
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- Peptide Models of Immunological Recognition: Paratope Dissection by Multiple Peptide Synthesis
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- Concepts for a Model of Good Medical Laboratory Services
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- IFCC-WorldLabNews
Articles in the same Issue
- New Trends in Immunoassay Methodology
- Progress in Immunoassay Technology
- Standardization of Steroid Immunoassays – In Theory an Easy Task
- Thermodynamic Parameters in Immunoassay
- Engineering of an Anti-Steroid Antibody: Amino Acid Substitutions Change Antibody Fine Specificity from Cortisol Estradiol
- Antigenic Definition of Cardiac Troponin I
- Peptide Models of Immunological Recognition: Paratope Dissection by Multiple Peptide Synthesis
- Remodeling of Glycoprotein and Carbohydrate Antigens
- Strongly Enhanced Serum Levels of Vascular Endothelial Growth Factor (VEGF) after Poly-trauma and Burn
- Rapid Electrophoretic Separation of Pre-ß-Migrating High Density Lipoproteins Using Automated PhastSystem(tm): Application to Analysis of Lecitihin: Cholesterol Acyltransferase-Deficient Plasma
- Decrease in High Density Lipoprotein Cholesterol (HDL-C) Levels Following Gemfibrozil Therapy
- Reference Intervals for Serum Cystatin C and Serum Creatinine in Adults
- Concepts for a Model of Good Medical Laboratory Services
- Diagnostic Kits Derived from Standard Method "DGKC 94" as a Potential Tool for Improvement of Analytical Standardization and Clinical Utility of Alkaline Phosphatase
- Antioxidant Interferences in Superoxide Dismutase Activity Methods Using Superoxide Radical as Substrate
- Cytokines and Soluble Adhesion Molecules in Laboratory Diagnostics. Report from a Meeting Held at the German Cancer Research Center in Heidelberg on 25 November 1997
- IFCC-WorldLabNews
