Here, we show the development of a pipeline for the robust characterization of nanoparticles prior-to and following introduction to protein containing media. We have addressed some of the key challenges facing nanoparticle-protein interactions as shown by our findings. These data suggest that surface chemistry plays a significant role in governing nanoparticle-protein interactions, where we demonstrated that the carboxylate-modified nanoparticles adsorbed the least amount of protein, accompanied with the lowest increase in particle size following incubation across all conditions (2-hour, 24-hour, 0.85 cm/s, and 8.5 cm/s). We show that amine-modified particles are positively charged before incubation (Table ), and adsorb more proteins onto the particle surface following incubation when compared to the negativelycharged carboxylate-modified polystyrene latex nanoparticles. This is the result of the incubation medium containing predominantly negatively-charged proteins at physiological pH . The unmodified polystyrene latex nanoparticles, despite having a similar charge to carboxylate-modified nanoparticles showed more protein adsorption when compared to carboxylate-modified nanoparticles and possessed a higher mean particle size. This is due to the exposed carboxyl groups found on the particle surface following functionalisation which impacts nanoparticle-protein interactions . Similar observations were made by Lundqvist et al. , which also compared polystyrene latex nanoparticles with similar surface chemistry incubated within human serum.