Establishment of substance P modified affinity chromatography for specific detection and enrichment of Mas-related G protein-coupled receptor X2.

Abstract Mas-related G protein-coupled receptor X2 (MrgX2) has been identified to be critical in drug-induced pseudo-allergic reactions and allergic diseases. Herein, an affinity high-performance liquid chromatography was established for the specific detection and enrichment of MrgX2. Substance P was used as an affinity ligand and immobilized on a glutaraldehyde-modified amino silica gel. The successful grafting of substance P was characterized by infrared spectroscopy, elemental analysis, X-ray photoelectron spectroscopy, thermogravimetric analysis, and nitrogen adsorption and desorption analyzes. The prepared materials were then used as the stationary phase to investigate the retention behavior of MrgX2 recombinant protein on the affinity column. The results obtained with the analytical techniques show the specificity and selectivity of the MrgX2 recombinant protein on the affinity column. The repeatability and reproducibility for the analysis of MrgX2 on the NH2-Silico@GD@SP column show relative standard deviation (RSD) values lower than the acceptance criteria of 2 and 5% of retention time, and RSD of peak areas