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ELISA binding assays are often used as fit-for-purpose potency assays at early stage of product development for monoclonal antibody and bispecific antibody products. However, the development of an ELISA can be very challenging for bispecific antibody given the complexity of the molecule including multiple binding targets and various binding kinetics and affinity. Our study has shown Surface Plasmon Resonance (SPR) technology plays a critical role to support the development of an ELISA for bispecific antibody by determining the binding kinetics, affinity, and avidity to various targets, and suggesting an optimal ELISA assay format. This strategy can also be applied to bioassay development of other complex modalities.