title: Development and validation of a novel SPR-based assay principle for bispecific molecules.
abstract: Surface plasmon resonance (SPR) is increasingly applied in drug discovery, early development and production. However, there are remarkably few reports describing the application of SPR in a regulated environment. Here, we describe a novel SPR-based assay, which enables us to assess the binding activity of a bivalent-bispecific anti-Ang-2/anti-VEGF antibody to both targets in a single setup. Validation of the assay revealed a high level of precision, accuracy, linearity and specificity. Upon analysis of temperature stressed samples it could be shown that firstly, the assay is able to indicate function-loss and secondly, it allows the parallel analysis of an additional interaction. Therefore, the described assay is highly suitable for quality assessment of the Ang-2/VEGF CrossMab. Additionally, the use of SPR in the context of assay development and routine use in a GMP environment is discussed.
authors.affiliation: Large Molecule Research, Pharma Research and Early Development, Roche Innovation Center Penzberg, Germany. Electronic address: christian.gassner@roche.com.
authors.forename: C
authors.initials: C
authors.lastname: Gassner
authors.affiliation: pRED Informatics, Pharma Research & Early Development, Roche Innovation Center Penzberg, Germany.
authors.forename: F
authors.initials: F
authors.lastname: Lipsmeier
authors.affiliation: Pharma Technical Development Europe (Biologics) Analytics, Hoffmann-La Roche Ltd., 4070 Basel, Switzerland.
authors.forename: P
authors.initials: P
authors.lastname: Metzger
authors.affiliation: Pharma Technical Development Europe (Biologics) Analytics, Hoffmann-La Roche Ltd., 4070 Basel, Switzerland.
authors.forename: H
authors.initials: H
authors.lastname: Beck
authors.affiliation: Pharma Technical Development Europe (Biologics) Analytics, Hoffmann-La Roche Ltd., 4070 Basel, Switzerland.
authors.forename: A
authors.initials: A
authors.lastname: Schnueriger
authors.affiliation: Large Molecule Research, Pharma Research and Early Development, Roche Innovation Center Penzberg, Germany.
authors.forename: J T
authors.initials: JT
authors.lastname: Regula
authors.affiliation: Large Molecule Research, Pharma Research and Early Development, Roche Innovation Center Penzberg, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Moelleken
mesh_terms: D042702:Angiopoietin-2 / Q000378:metabolism*; D018033:Antibodies, Bispecific / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D018448:Models, Immunological; D020349:Surface Plasmon Resonance; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
chemical_list: D042702:Angiopoietin-2; D018033:Antibodies, Bispecific; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A
keywords: Activity assessment; Binding assay; Bispecific antibody; Bridging assay; Surface plasmon resonance