Particle-based epitope analysis of a monoclonal antibody
- A method for the examination of the epitope of a monoclonal antibody against a peptide related to hepcidin-25, known as a biomarker in the ion metabolism, was developed. Characterizing the exact binding site helps to understand the properties of the antibody. A peptide consists of different amino acids, each contributing differently to the binding strength. To examine the influence of the amino acids, each amino acid was subsequently exchanged by glycine. If the respective amino acid is critical for binding to the antibody, a significant drop in the binding affinity should be observed. The epitope screening of monoclonal antibodies is a complex and elaborate procedure. For this approach, variants of the peptides were synthesized by conventional Fmoc-based peptide synthesis. The solid support was chosen to be suitable for the synthesis and additional screening against the antibody. To determine the binding affinity, the bead-bound peptides were incubated with a fluorophore-labeledA method for the examination of the epitope of a monoclonal antibody against a peptide related to hepcidin-25, known as a biomarker in the ion metabolism, was developed. Characterizing the exact binding site helps to understand the properties of the antibody. A peptide consists of different amino acids, each contributing differently to the binding strength. To examine the influence of the amino acids, each amino acid was subsequently exchanged by glycine. If the respective amino acid is critical for binding to the antibody, a significant drop in the binding affinity should be observed. The epitope screening of monoclonal antibodies is a complex and elaborate procedure. For this approach, variants of the peptides were synthesized by conventional Fmoc-based peptide synthesis. The solid support was chosen to be suitable for the synthesis and additional screening against the antibody. To determine the binding affinity, the bead-bound peptides were incubated with a fluorophore-labeled antibody. After the incubation, the beads were placed on a slide and the fluorescence was detected by a fluorescence scanner. A high fluorescence indicates a high binding affinity and vice versa.…
Autor*innen: | Timm Schwaar |
---|---|
Koautor*innen: | J. W. van den Berg, Ioana M. AbbasORCiD, Michael G. WellerORCiD |
Dokumenttyp: | Posterpräsentation |
Veröffentlichungsform: | Präsentation |
Sprache: | Englisch |
Jahr der Erstveröffentlichung: | 2017 |
DDC-Klassifikation: | Naturwissenschaften und Mathematik / Chemie / Analytische Chemie |
Freie Schlagwörter: | Antibody; Epitope analysis |
Veranstaltung: | Tag der Chemie 2017 |
Veranstaltungsort: | Berlin, Germany |
Beginndatum der Veranstaltung: | 05.07.2017 |
Enddatum der Veranstaltung: | 05.07.2017 |
Verfügbarkeit des Dokuments: | Datei im Netzwerk der BAM verfügbar ("Closed Access") |
Datum der Freischaltung: | 02.08.2017 |
Referierte Publikation: | Nein |