Controlled synthesis of open-mouthed epitope-imprinted polymer nanocapsules with a PEGylated nanocore and their application for fluorescence detection of target protein

Abstract

Epitope imprinting is an effective way to create artificial receptors for protein recognition. Surface imprinting with immobilized templates and sacrificial supports can generate high-quality imprinted cavities of homogeneous orientation and good accessibility, but it is still challenging to fabricate nanoscale imprinted materials by this approach. Herein, we propose a method for the controlled synthesis of open-mouthed epitope-imprinted polymer nanocapsules (OM-MIP NCs) by limiting the imprinting polymerization on the template-bearing side of the Janus nanoparticles (JNPs). Concurrent bromoacetyl (Ac–Br) and 2-bromoisobutyryl (iB–Br) functionalization of the major portion of SiO₂ nanoparticles is achieved via the molten-wax-in-water Pickering emulsion approach. The cysteinyl-derived epitope templates are immobilized through the Ac–Br groups, and then surface imprinting is fulfilled via ATRP initiated by the iB–Br groups. The SiO₂ supports are partially etched and then PEGlated, affording OM-MIP NCs with a PEGylated nanocore. The inside nanocore can facilitate collection of the NCs by centrifugation, and its PEGylation can inhibit non-specific binding. The surface imprinting can be optimized through the ATRP time, and the etching can be tailored via the concentration of NH₄HF₂ employed. For proof-of-concept, with a C-terminus nonapeptide of bovine serum albumin (BSA) chosen as a model epitope and polymerizable carbon dots added to the pre-polymerization solution, fluorescent OM-MIP NCs were fabricated for BSA sensing. The as-synthesized NCs exhibited satisfactory detection performance, with an imprinting factor of 6.1, a limit of detection of 38.1 nM, a linear range of 0.25–6 μM, and recoveries of 98.0 to 104.0% in bovine serum samples.