Tailoring molecular recognition in predesigned multifunctional enzyme mimicking porphyrin imprinted interface for high affinity and differential selectivity; sensing etoposide in lung cancer patients

Abstract

Nanozymes are cost-effective and robust but they lack specificity and selectivity, limiting their potential practical applications. Herein, molecularly imprinted polymers (MIPs) were grown in combination with multifunctional 5,10,15,20-tetrakis(4-hydroxyphenyl)-21H,23H-porphyrin (THPP) oxidase-like nanozyme to engineer THPP@MIP interface with high affinities and differential selectivity for structurally related target analytes. THPP nanozyme displayed a high level of predefined binding affinity for etoposide (ETO), and served as a predesigned functional monomer to rationally tailor the selectivity of THPP@MIP surface in the presence of different guest molecules. THPP nanozyme in combination with conventional monomers was imprinted on a portable and disposable cellulose paper matrix under UV light to create a UV-cured imprinted interface for optical detection of ETO. The THPP@MIP enzyme mimicking interface, having ETO specific and selective target recognition pockets, catalyzed the oxidation of colorless 3,3′,5,5′-tetramethylbenzidine (TMB) to generate visible blue oxidized TMB (oxTMB) without exogenous hydrogen peroxide (H2O2). The ETO binding on the THPP@MIP surface blocked the channels for TMB access to THPP cavities. The THPP@MIP sensor permitted to detect ETO in the linear range of 0.005–10 μg mL−1, with a limit of detection (LoD) of 0.002 μg mL−1, and showed a remarkable specificity and selectivity against other drug molecules. Furthermore, the THPP@MIP sensor successfully differentiated the serum samples of lung cancer patients and healthy volunteers. The obtained results were validated with standard High performance liquid chromatography-mass spectrometry (HPLC/MS) analysis of the serum samples. Additionally, ETO injection/infusion solutions and ETO-free serum samples were used to perform the matrix effect and recovery studies. This work demonstrates that molecular imprinting with predesigned, enzyme mimicking, high-affinity functional monomer can serve as a highly selective and specific universal interface for broad spectrum sensing applications in various analytical domains.