[DOT 2020] Identify Effective Therapeutics with Simple, Quantitative Cell-Based Assays That Measure Receptor Internalization

[DOT 2020] Identify Effective Therapeutics with Simple, Quantitative Cell-Based Assays That Measure Receptor Internalization
File Name/Number:
DOT 2020

Year:
2020


Both multi- and single-pass membrane receptors constitutively exhibit complicated and dynamic membrane receptor trafficking as well as internalization induced by ligand or compound binding. Measuring how therapeutic molecule binding to membrane receptors affects receptor internalization can provide insight into drug tolerance, unwanted side effects, diseases, and ultimately help with identifying safer drugs. Additionally, antibody-based therapeutics that target specific receptors such as the immuno-oncology checkpoint receptors can induce high levels of internalization and serve as candidates for conjugation to cytotoxic compounds. These therapeutic Antibody-Drug Conjugates (ADCs) take advantage of both antibody specificity for tumor antigens and antibody induction of receptor internalization to deliver cytotoxic drugs to cancer cells effectively. We present here a collection of cell-based assays that use the industry-validated Enzyme Fragment Complementation (EFC) technology to monitor receptor internalization activated by small molecules or biologics. These assays are easy-to-use and offer a quantitative means of measuring receptor internalization that is amenable to high-throughput screening. PathHunter® Internalization Assays provide an effective alternative to more cumbersome, less quantitative techniques using imaging, antibodies or dyes/labels to determine internalization. Assays for both single-pass membrane receptors such as receptor tyrosine kinases (RTK) and checkpoint receptors and multi-pass membrane receptors like GPCRs have been developed using this generic platform. An additional assay format for GPCRs specifically measures activated GPCR internalization using β-arrestin recruitment. Results from such membrane receptor internalization assays can provide important safety and efficacy information during drug development.

Listen to a short recording of the poster by Jennifer Lin-Jones, Ph.D.