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Updated: Jan 25, 2026

Nano-Differential Scanning Fluorimetry for Screening in Fragment-based Lead Discovery
Published on: May 16, 2021
Site-Specific Immobilization of β2-AR Using O6-Benzylguanine Derivative-Functionalized Supporter for High-Throughput
Jing Wang1, Yuxin Wang1, Jiajun Liu1
1Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Sciences , Northwest University , Xi'an , Shaanxi 710069 , China.
Abstract:
The past decade has witnessed the great promise of strategies for ligand discovery based on surface-immobilized GPCRs. We present here a method for preparation of immobilized GPCRs. Key features include covalent immobilization with high specificity and robust application in drug-receptor interaction analysis and ligand screening. In our example assay using beta2-adrenergic receptor (β2-AR), the human DNA repair protein O6-alkylguanine-DNA alkyltransferase (hAGT) fusion receptor expressed in Escherichia coli was directly captured onto polyethylene glycol polyacrylamide (PEGA) resin. We observed even distribution and physiological functions of β2-AR on the resin. The immobilized β2-AR as a stationary phase enabled us to rapidly determine the binding of four drugs to β2-AR. By coupling this assay to mass spectrometry, we screened rosmarinic acid as a bioactive compound targeting β2-AR in Fructus Perillae. We concluded that O6-benzylguanine derivative-functionalized supporter is promising for specific immobilization of hAGT-tagged proteins; immobilized receptor chromatography has great potential in screening receptor-binding leads from herbal plants or traditional medicine recipes.
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