With the rapid developments of nanotechnology, chances of exposing nanoscale particles to humans (e.g., workers and consumers) also increase correspondingly, which raises serious concerns on their biosafety. Entrance of nanoparticles into diverse biological environment endows them with new and dynamic biological identities as the so-called nanoparticle–protein corona. Therefore, understanding the role of these nanoparticle–protein coronas and resulting biological responses is crucial, as it helps to clarify the biological mechanism and prevent the potential adverse effects of nanoparticles. In this review, we summarize the latest developments relating to the nanoparticle–protein interaction and corresponding biological responses, with an emphasis on the characterization methods, induced biological effects and possible molecular mechanisms. In addition, we overview both the challenges and opportunities (particularly in nanomedicine) raised by this entrance of nanoparticles into the living creatures, especially human beings, with some future perspectives based on our understanding.