Nonspecific luminometric assay for monitoring protein adsorption efficiency and coverage on nanoparticles

摘要: Nonspecific assays utilizing time-resolved luminescence resonance energy transfer (TR-LRET) are developed for two applications: to monitor protein adsorption efficiency and assess the degree of surface coverage on solid phase. We successfully measure non-sedimenting nanoparticles since this has been notoriously difficult determine. Monitoring does not require with adsorbed be washed it is based competitive between non-adsorbed analyte acceptor-labeled donor europium(III) polystyrene nanoparticles. The application assessing performed final coated requires no fluorescent labeling studied protein. This utilizes partly covered methods detect either or uncovered nanoparticle surface, proteins achievable traditional total quantification applied monitoring both detected their separation required. Thus, user-friendly, centrifugal instrumentation, applicable also small requiring ultracentrifugation. No special expertise investment in costly instruments required compared existing methods, such as spectroscopic techniques, isothermal titration calorimetry, plasmon resonance. a reference literature method that comprised analysis fluorescently labeled saturation reached at similar concentrations showing reliability assay. Our results suggest applications could exploited rapid tools studies quality control characterization coating processes.