The nanoparticle corona—a molecular layer adsorbed on nanoparticle surfaces—plays a crucial role in controlling molecular interactions and enabling applications in catalysis, nanoparticle separations, and sensing technologies. It underpins Corona Phase Molecular Recognition (CoPhMoRe), where the adsorbed layer adopts a conformation that selectively binds a specific molecular configuration. While tailoring the corona has enabled the molecular recognition of diverse analytes, quantitatively characterizing its properties remains challenging. Herein, we advance Molecular Probe Adsorption (MPA), applying it across a broad range of data sets to further validate and refine the technique. MPA utilizes a fluorescent probe quenched upon adsorption to quantify solvent-exposed surface area via adsorption isotherms. We leverage MPA to elucidate recognition mechanisms in five corona phases and expand the library of characterized CoPhMoRe constructs by 20, enabling comprehensive comparative analyses. Additionally, we examine how polymer stiffness, quantified by persistence length, influences corona formation on single-walled carbon nanotubes (SWCNTs). Our results reveal that as the calculated polymer persistence length increases from 1.16 nm to ~4 nm, the q/KD value, a proxy for exposed surface area, rises from ~200 M
-1 to 1,600 M
-1. In agreement with theoretical predictions, we find that sufficiently flexible polymers achieve greater SWCNT surface coverage, offering new insights for optimizing polymer-based corona phases. Furthermore, we establish structure–property relationships linking MPA-derived surface area to adsorption parameters, demonstrating that ssDNA and high molecular weight polymers exhibit differing probe–corona interactions, with binding affinities varying by a factor of ~2.7. Lastly, we show that MPA can be integrated with molecular dynamics simulations and thermodynamic modeling to predict analyte binding affinities for 42 phytohormones, enabling computational CoPhMoRe screening. This integration paves the way for rational sensor design without extensive experimental screening. Our findings underscore the utility of MPA in advancing nanomaterial-based sensing technologies through quantitative corona characterization and provide a framework for the rational design of selective nanosensors.
Keywords: molecular probe adsorption, nanoparticle, surface area, molecular recognition
