Conjugation effects on antibody-drug conjugates: evaluation of interaction kinetics in real time on living cells.

Abstract

Antibody-drug conjugates (ADC) have shown promising effects in cancer therapy by combining the target specificity of an antibody with the toxicity of a chemotherapeutic drug. As the number of therapeutic antibodies is significantly larger than those used as ADCs, there is unused potential for more effective therapies. However, the conjugation of an additional molecule to an antibody may affect the interaction with its target, altering association rate, dissociation rate, or both. Any changes of the binding kinetics can have subsequent effects on the efficacy of the ADCs, thus the kinetics are important to monitor during ADC development and production. This paper describes a method for the analysis of conjugation effects on antibody binding to its antigen, using the instrument LigandTracer and a fluorescent monovalent anti-IgG binder denoted FIBA, which did not affect the interaction. All measurements were done in real time using living cells which naturally expressed the antigens. With this method the binding profiles of different conjugations of the therapeutic anti-EGFR antibody cetuximab and the anti-CD44v6 antibody fragment AbD15171 were evaluated and compared. Even comparatively small modifications of cetuximab altered the interaction with the epidermal growth factor receptor (EGFR). In contrast, no impact on the AbD15171-CD44v6 interaction was observed upon conjugation. This illustrates the importance to study the binding profile for each ADC combination, as it is difficult to draw any general conclusion about conjugation effects. The modification of interaction kinetics through conjugation opens up new possibilities when optimizing an antibody or an ADC, since the conjugations can be used to create a binding profile more apt for a specific clinical need.

• Authors: Sina Bondza, Jonas Stenberg, Marika Nestor, Karl Andersson, Hanna Björkelund
• Journal: Molecular pharmaceutics (Mol Pharm) Vol. 11 Issue 11 Pg. 4154-63 (Nov 03 2014) ISSN: 1543-8392 [Electronic] United States
• PMID: 25252166 (Publication Type: Evaluation Study, Journal Article)

Chemical References

• Antibodies, Anti-Idiotypic
• Antibodies, Monoclonal
• Antibodies, Monoclonal, Humanized
• CD44v6 antigen
• Fluorescent Dyes
• Hyaluronan Receptors
• Immunoconjugates
• anti-IgG
• EGFR protein, human
• ErbB Receptors
• Cetuximab

Topics

• Antibodies, Anti-Idiotypic (immunology, metabolism)
• Antibodies, Monoclonal (immunology, metabolism)
• Antibodies, Monoclonal, Humanized (immunology, metabolism)
• Carcinoma, Squamous Cell (drug therapy, metabolism, pathology)
• Cetuximab
• ErbB Receptors (immunology, metabolism)
• Fluorescent Dyes
• Humans
• Hyaluronan Receptors (immunology, metabolism)
• Immunoconjugates (chemistry)
• Kinetics
• Protein Interaction Domains and Motifs (drug effects)
• Tumor Cells, Cultured