Nanoparticles (NPs) with unique physicochemical properties possess distinct properties that are ultimately superior to that of the properties of their bulk counterparts. Hence, their market for industrial applications has counted up to the maximum. Nanoparticle embedded cosmetics, balls, fabrics, water filters, etc. has already occupied their significant part in the commercial sector. Such nanoparticle incorporated products and nano based therapeutics provides enough opportunity for the exposure of nanoparticles to that of human beings from nature either out of pollution or via their biomedical applications. Inhalation or the intentional administration of nano based products into human being results out into the nanoparticle-protein corona that determines the fate of nanoparticle internalization into the body. Such kinds of interaction occurring at the molecular level not only determine the fate of NPs, but also the binding accompanied conformational changes in biomolecules. Also, NPs unique higher surface area to volume ratio promotes the biomolecules aggregation, defined as the protein fibrillation at molecular level. Several spectroscopic techniques occupy prominent position for studying the above phenomena in a much simplified manner. On accounting the above physico-chemical and biological behavioral properties of NPs, the understanding of the impact of nanoparticle on the protein conformation and fibrillation upon binding should be reviewed enough to understand the biocompatibility of nanoparticles for futuristic biomedical applications. Hence, this review encompasses the brief definition and the relationship between nanoparticle, protein corona, and fibrillation process with that of the biophysical methods. Copyright © 2016 American Scientific Publishers.