Dual-Color Bioluminescent Sensor Proteins for TherapeuticDrug Monitoring of Antitumor Antibodies

摘要

Monitoring the levels of therapeutic antibodies in individual patients would allow patient-specific dose optimization, with the potential for major therapeutic and financial benefits. Our group recently developed a new platform of bioluminescent sensor proteins (LUMABS; LUMinescent AntiBody Sensor) that allow antibody detection directly in blood plasma. In this study, we targeted four clinically important therapeutic antibodies, the Her2-receptor targeting trastuzumab, the anti-CD20 antibodies rituximab and obinutuzumab, and the EGFR-blocking cetuximab. A strong correlation was found between the affinity of the antibody binding peptide and sensor performance. LUMABS sensors with physiologically relevant affinities and decent sensor responses were obtained for trastuzumab and cetuximab using mimotope and meditope peptides, respectively, with affinities in the 10^–7 M range. The lower affinity of the CD20-derived cyclic peptide employed in the anti-CD20 LUMABS sensor (Kd = 10^–5 M), translated in a LUMABS sensor with a strongly attenuated sensor response.The trastuzumab and cetuximab sensors were further characterized withrespect to binding kinetics and their performance in undiluted bloodplasma. For both antibodies, LUMABS-based detection directly in plasmacompared well to the analytical performance of commercial ELISA kits.Besides identifying important design parameters for the developmentof new LUMABS sensors, this work demonstrates the potential of theLUMABS platform for point-of-care detection of therapeutic antibodies.