Construction and application of covalently bonded CD147 cell membrane chromatography model based on polystyrene microspheres.

Abstract

In this study, a novel cell membrane chromatography (CMC) model was developed to investigate cluster of differentiation 147 (CD147) targeted anti-tumor drug leads for specific screening and ligand-receptor interaction analysis by SNAP-tagged CD147 fusion protein conjugation and polystyrene microspheres (PS) modification. Traditional Chinese medicines (TCMs) are widely used in the treatment of cancer. CD147 plays important roles in tumor progression and acts as an attractive target for therapeutic intervention; therapeutic drugs for CD147-related cancers are limited to date. Thus, ascreening method for active components in TCMs is crucial for the further research and development of CD147 antagonists. However, improvement is still needed to perform specific and accurate drug lead screening using the CMC-based method. Recently, our group developed acovalently immobilized receptor-SNAP-tag/CMC model using silica gel as carrier. Besides thecarboxyl group on multi-step modified silica particles, theamino group of benzyl-guanine (BG, substrate of SNAP-tag) also possesses reactivity towards thecarboxyl group on available carboxyl-modified PS. Herein, we used PS as carrier and anextended SNAP-tag with CD147 receptor to construct the PS-BG-CD147/CMC model for active compound investigation coupled with HPLC/MS and applied this coupled PS-BG-CD147/CMC-HPLC/MS two-dimensional system to drug lead screening from Nelumbinis Plumula extract (NPE) sample. In addition, to comprehensively verify the pharmacological effects of screened ingredients, acell proliferation inhibition assay was performed, and the interaction between the ingredients and CD147 was studied by thefrontal analysis method. This study developed a high-throughput PS-based CMC screening platform, which could be widely applied and utilized in chromatographic separation and drug lead discovery.