1. SPR-based assays enable the full functional analysis of bispecific molecules
- Author
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Jörg T. Regula, W. Meschendoerfer, Florian Lipsmeier, Jörg Moelleken, and Christian Gassner
- Subjects
Vascular Endothelial Growth Factor A ,0301 basic medicine ,Bispecific antibody ,Clinical Biochemistry ,Activity assessment ,Pharmaceutical Science ,Nanotechnology ,Biosensing Techniques ,Computational biology ,Ligands ,01 natural sciences ,Analytical Chemistry ,Angiopoietin-2 ,03 medical and health sciences ,Antigen ,Reference Values ,Antibodies, Bispecific ,Drug Discovery ,Humans ,Molecule ,Surface plasmon resonance ,Immunoglobulin Fragments ,Spectroscopy ,Functional analysis ,Chemistry ,010401 analytical chemistry ,Reproducibility of Results ,Surface Plasmon Resonance ,Fusion protein ,0104 chemical sciences ,030104 developmental biology ,Drug development ,Drug Design ,Linear Models ,Biological Assay ,Protein Binding - Abstract
The increasing complexity of novel biotherapeutics such as bispecific antibodies or fusion proteins raises new challenges for functional characterization. When compared to standard antibodies, two individual interactions and the inter-dependency of binding events need to be considered for bispecific antibodies. We have previously described an SPR-based assay setup, which enables us to assess the binding activity of a bivalent-bispecific molecule to both targets simultaneously and - in addition to one individual target - in a single setup. However, there might be some pitfalls when applying the bridging assay, e.g. change of antigen activity upon immobilization. Therefore, we have developed an alternative SPR-based assay principle, which allows the individual assessment of both targets in solution. Comparison of data between the assays showed that simultaneous binding can be calculated based on both individual readouts, and revealed a good correlation. Hence, both SPR-based assay principles allow a "full" functional analysis of a bispecific CrossMab in only one assay. The assay principles can be qualified and enable an efficient drug development.
- Published
- 2017