Bispecific biotherapeutics present a unique bioanalytical challenge for drug development. NAb assay development for a bispecific therapeutic encountered significant challenges derived from a low-affinity drug target binding domain and a high drug trough serum concentration. These obstacles were overcome largely by implementing a homogenous multiplex bead-based competitive ligand binding assay based on the AlphaLISA(TM) . The duplex NAb assay was designed to enable simultaneous measurement of NAb responses to both functional domains of the bispecific biotherapeutic. An Affinity Capture Elution (ACE) method was utilized to improve drug tolerance by isolating NAbs from serum matrix prior to the assay. The homogenous, no-wash nature of the bead-based detection method, allowed the detection of the low-affinity drug-target interaction in the NAb assay. Interestingly, the novel homogeneous NAb assay platform combined with ACE-based NAb purification supported sensitive NAb detection amidst milligram level of circulating drug concentrations after method optimization.
Learning Objectives:
Upon completion, participants will have a better understanding of the Alphalisa technology platform.
Upon completion, participants will understand how I applied the Alphalisa technology platform to a bispecific for NAb detection.
Upon completion, participants will understand the challenges that were overcome when developing a bispecific NAb assay.