Development of a Stationary Phase of Vascular Smooth Muscle Cell Membrane Chromatography and Its Chromatographic Affinity Characteristics

A novel stationary phase of vascular smooth muscle cell membrane chromatography (VSM-CMC) was developed by immobilizing the vascular smooth muscle cell membrane onto the surface of silica and was presented for bioaffinity chromatography. The protein level and Na+, K+-ATP enzymatic activity of the vascular smooth muscle cell membrane stationary phase (VSM-CMSP) were detected. The surface characteristics of the VSM-CMSP were tested using scanning electron microscopy and surface energy spectrometry. The retention characteristics of four dihydropyridines (amlodipine besylate, nicardipine hydrochloride, nitrendipine and nifedipine) were investigated using a VSM-CMSP column (10 mm × 2 mm, I.D.) packed with VSM-CMSP. The logarithm of the capacity factor (logk′) was taken as a measure of the affinities of the calcium antagonists toward the vascular smooth muscle cell membrane and receptors. The surface characteristics of the VSM-CMSP were very different from that of the normal and reversed stationary phase, and the VSM-CMSP was found to have cell membrane activity and chromatographic separation. Moreover, there was a significant correlation between the affinity in the VSM-CMC system and the effect in vitro with respect to the pharmacological effect. It is concluded that the VSM-CMC system can serve as a type of bioaffinity chromatography for studying the interaction between drug molecules and target sites (e.g., receptors) in cell membranes, and for screening active compounds from complex agents.