An image-based Abplex method for high-throughput GPCRs antibody discovery

As the field of antibody therapeutics advances rapidly, membrane proteins, particularly G protein-coupled receptors (GPCRs), have emerged as highly sought-after drug targets. However, the challenges associated with extracting membrane proteins have created a demand for effective antibody screening systems targeting these proteins. In this study, we propose developing an innovative antibody screening strategy (Abplex) based on high-content imaging. This approach leverages intact cells that express target membrane proteins, facilitating the presentation of proteins in their native conformation. Furthermore, it acquires both specific and non-specific binding signals in a single well, thereby bolstering the robustness of the outcomes. The technique involves just one step and can be completed within 50 min, enabling the analysis of a single sample in just one second. The amalgamation of dependable experimental findings, a simplified workflow, reduced hands-on time, and a swift analytical pace positions our method for superior throughput and precision when juxtaposed with traditional techniques such as CbELISA and FACS. Moreover, we introduce the concept of cell barcoding, wherein cells are labeled with different fluorescence spatial patterns. This feature allows for multiplexed detection to meet the needs of various experiments. The characteristics of Abplex promise to expedite GPCR-targeting antibody discovery, advance therapeutics and enable new disease treatments. The challenges associated with extracting membrane proteins have created a demand for effective antibody screening systems targeting GPCRs. In this study, we propose developing an innovative antibody screening strategy (Abplex) based on high-content imaging. The amalgamation of dependable experimental findings, a simplified workflow, reduced hands-on time, and swift analytical pace positions our method for superior throughput and precision when juxtaposed with traditional techniques. Abplex is versatile as it can be used to detect cross-species reactivity at the same time as antibody binding screening. Specific, non-specific, and cross-species binding signals can be investigated simultaneously in one well. The high-throughput Abplex method featured with barcoded cells contributes to the GPCRs antibody discovery.image