Comparative Study on Selective Profiling of Cardiovascular and Cerebrovascular Disease-Associated Proteins Using Two Statin-Based Magnetic Separation Materials

Abstract

This study established a drug-associated protein screening strategy leveraging the principle of specific protein-drug interactions. Two first-line lipid-lowering drugs, atorvastatin (AN) and rosuvastatin (RSV), were covalently immobilized onto nanoparticle surface to fabricate novel magnetic separation materials, Fe₃O₄@AN and Fe₃O₄@RSV. Initial validation of the strategy was performed using fetal bovine serum (FBS) as a model sample, with alterations in protein amino acid sequence coverage serving as the evaluation metric. Fe₃O₄@AN speicificly adsorbed four proteins from FBS, all of which were implicated in cardiovascular diseases (CVDs). Similarly, Fe₃O₄@RSV enriched five CVD-associated proteins from FBS. Three proteins (Q3T052, P1276, and Q58D62) were co-enriched by both materials. Subsequently, the strategy was applied to clinical serum samples from CVD patients and heathy controls to screen disease-relevant proteins via label-free quantitative proteomics. Comparative analysis with healthy controls revealed that Fe₃O₄@AN and Fe₃O₄@RSV selectively enriched 23 and 37 differentially expressed proteins (DEPs) from sera of CVD patients, respectively. Gene Ontology (GO) and pathway enrichment analyses indicated distinct functional pathways for proteins captured by each material, and both sets were significantly associated with CVD-related biological processes and cellular components. Notably, six overlapping DEPs were co-identified by the two materials, all demonstrating critical roles in CVD pathogenesis. Among these, two proteins (P02042, P14174) were exclusively detected in patient sera, while Q14624 was detectable in CVD patient serum, healthy human serum, and FBS serum but significantly upregulated in CVD patient serum. Collectively, this protein-drug interaction-based screening strategy exhibits broad applicability and establishes a novel paradigm for rational drug design and development.