Enhanced Stability Designs of Cell Membrane Chromatography for Screening Drug Leads

Abstract

Cell membrane chromatography (CMC) is an effective method to screen bioactive components acting on specific receptor from complex systems. CMC keeps the biological activity of the membrane receptors and improves the efficiency of screening. However, the traditional CMC suffered from poor stability, resulting in limited life span and low reproducibility. Which greatly limits the application of this method, especially limit the application of precious cell materials. To solve this problem, cyanuric chloride (TCT)-functionalized silica gel was used for covalent immobilization of cell membranes. TCT reacts with amino groups on the cell membrane to form covalent bonds. In this way, the cell membranes are not easy to fall off. The column life of the TCT-decorated epidermal growth factor receptor (EGFR)/CMC column was extended to more than 8 days, while the normal CMC column of the control group dropped sharply in the first 3 days. TCT-decorated EGFR/CMC online HPLC-IT-TOF-MSn system was successfully applied for potential screening of drug leads from Trifolium pratense L.. One potential drug lead acts on the EGFR receptor as formononetin was screened. Such immobilization method owing to the properties of mild conditions, fast reaction speed, high yield and small loss of membrane protein activity. In a word, our strategy of covalent immobilization of cell membrane receptors improves the stability of CMC.