DOI: https://doi.org/10.29363/nanoge.cybioel.2024.052
Publication date: 28th June 2024
G protein-coupled receptors (GPCRs) constitute the largest family of membrane receptors [1]. Their abundance and involvement in numerous physiological processes within the human body, render GPCRs pivotal targets for pharmaceutical intervention, with over one-third of FDA-approved drugs targeting these receptors [2]. While assessing binding characteristics of GPCRs is essential during drug development, common optical assays rely on ligand labelling, which impacts binding characteristics and increases costs [3]. A cost-efficient alternative that forgoes labelling is presented by biomembrane-based bioelectronic sensing techniques, which employ cell-derived membrane models such as native supported lipid bilayers (SLBs) [4].
This work explores the feasibility of investigating GPCR ligand binding within cell-derived membrane models in the form of vesicles (blebs) and SLBs. Blebs and SLBs obtained from two distinct GPCR-expressing cell lines were probed via Nanoluciferase-based bioluminescence resonance energy transfer (NanoBRET) binding assays for GPCR functionality. It was observed that cell-derived vesicles could serve as viable substitutes for intact cells in discerning specific GPCR ligand binding characteristics. Additionally, these vesicles can be used to form native supported lipid bilayers and record characteristic binding behaviour. This means that functional GPCRs can be obtained via a simple chemically induced cell blebbing process and incorporated into native SLBs, representing a meaningful stride towards an electrical detection of GPCR ligand binding.
The integration of such biomembrane-based bioelectronic sensing techniques with microfabricated electronic device arrays presents a promising opportunity for high-throughput screening, with substantial potential to transform drug discovery. Future work will focus on complementary techniques to validate functionality of GPCRs in native SLBs formed from blebs as well as coupling biomembranes to electronic chips to explore the feasibility of electrically investigating GPCR ligand binding.
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