Understanding misfolding and aggregation of proteins is important in disciplines of proteins, pharmaceutical, and biomedical fields. Protein misfolding and assembly results in formation of amyloid fibrils leading to crippling conditions referred to as "amyloidosis”[1,2]. A combination of calorimetry and spectroscopy has been used to obtain mechanistic insights into effect of pluronics on the protein aggregation/ fibrillation. Isothermal titration calorimetry results suggest that interaction of protein with F68 is exothermic while with F127 it is endothermic due to more hydrophobicity of the latter pluronic. The conformation of protein fibrils formation was examined by TEM images and ThT binding assay, which demonstrated that pluronics promoted fibrillation process rather than inhibiting it. The pre- and post-micellar concentrations of pluronics on interaction with protein (at varying fibrillation stages) exhibit a reduction in enthalpy change value as the incubation time increases. This indicates the formation of amorphous aggregates due to which endothermic enthalpy is observed. AGGRESCAN program was used to identify aggregation-prone regions of protein. According to analysis, pluronics interact with the areas that are not prone to aggregation, exposing hotspot areas to the solvent and triggering formation of fibrils. Consequently, the potential use of these formed aggregates for drug delivery was explored. Interaction of anticancer drugs (5-fluorouracil and cytarabine) with the protein fibril systems was studied using calorimetry and spectroscopy. The results suggest one less order of binding for 5-fluorouracil with protein fibril systems than with cytarabine. When cytarabine is used with protein fibrils, the hydrophobic interaction predominates, according to endothermic enthalpy of interaction, but the electrostatic interaction predominates with 5-flourouracil. On the other hand, the former drug exhibited more adsorption on the surface of protein fibrils compared to the latter. Therefore, it is concluded that cytarabine has weak adsorption on fibril surfaces and is readily desorbed in cells, whereas 5-fluorouracil has relatively strong adsorption; as a result, the complex of LFF127 and 5-FU is fatal to malignant cells. These results are beneficial for exploring numerous applications of the formed protein fibrils/aggregates and in providing suitable strategies for the design and development of drug delivery agents.