Elucidating receptor occupancy (RO) of monoclonal antibodies (mAbs) is a crucial step in characterizing the therapeutic efficacy of mAbs. However, the in vivo assessment of RO, particularly within peripheral tissues, is greatly limited by current technologies. In the present study, we developed a bioluminescence resonance energy transfer (BRET)-based system that leverages the large signal:noise ratio and stringent energy donor-acceptor distance dependency to measure antibody RO in a highly selective and temporal fashion. This versatile and minimally invasive system enables longitudinal monitoring of the in vivo antibody-receptor engagement over several days. As a proof of principle, we quantified cetuximab-epidermal growth factor receptor binding kinetics using this system and assessed cetuximab RO in a tumor xenograft model. Incomplete ROs were observed, even at a supratherapeutic dose of 50 mg/kg, indicating that fractional target accessibility is achieved. The BRET-based imaging approach enables quantification of antibody in vivo RO and provides critical information required to optimize therapeutic mAb efficacy.
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http://dx.doi.org/10.1016/j.isci.2019.05.003 | DOI Listing |
ACS Sens
January 2025
Materials Interfaces Center, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, P. R. China.
Over recent years, the LUMinescent AntiBody Sensor (LUMABS) system, utilizing bioluminescence resonance energy transfer (BRET), has emerged as a highly effective method for antibody detection. This system incorporates NanoLuc (Nluc) as the donor and fluorescent protein (FP) as the acceptor. However, the limited Stokes shift of FP poses a challenge, as it leads to significant spectral cross-talk between the excitation and emission spectra.
View Article and Find Full Text PDFJ Enzyme Inhib Med Chem
December 2025
Laboratory of Biochemistry and Molecular Biology, Department of Chemistry, Life Sciences and Environmental Sustainability, University of Parma, Parma, Italy.
Antibiotic resistance stands as the foremost post-pandemic threat to public health. The urgent need for new, effective antibacterial treatments is evident. Protein-protein interactions (PPIs), owing to their pivotal role in microbial physiology, emerge as novel and attractive targets.
View Article and Find Full Text PDFBiosensors (Basel)
November 2024
Department of Pathology and Molecular Medicine, Queen's University, Kingston, ON K7L 3N6, Canada.
Recent advances in drug discovery have established biosensors as indispensable tools, particularly valued for their precision, sensitivity, and real-time monitoring capabilities. The review begins with a brief overview of cancer drug discovery, underscoring the pivotal role of biosensors in advancing cancer research. Various types of biosensors employed in cancer drug discovery are then explored, with particular emphasis on fluorescence- and bioluminescence-based technologies such as FRET, TR-FRET, BRET, NanoBRET, and NanoBiT.
View Article and Find Full Text PDFbioRxiv
December 2024
Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, La Jolla, CA, USA.
The constitutive (ligand-independent) signaling of G protein-coupled receptors (GPCRs) is being increasingly appreciated as an integral aspect of their function; however, it can be technically hard to detect for poorly characterized, e.g. orphan, receptors of the cAMP-inhibitory Gi-coupled (GiPCR) family.
View Article and Find Full Text PDFExtracell Vesicles Circ Nucl Acids
December 2023
School of Bioscience and Technology, Weifang Medical University, Weifang 261053, Shandong, China.
Cells have the capability to discharge extracellular vesicles (EVs) into a range of bodily fluids. Extracellular vesicles (EVs) encapsulate biological molecules such as proteins and nucleic acids, playing a role in facilitating cell-cell communication. They actively engage in a myriad of physiological and pathological processes.
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