Optimized Amyloid Nanofibrils for Lipase Immobilization in Targeted PDAC Nanotherapy

This study focuses on the design and optimization of amyloid-based nanofibril scaffolds derived from bovine serum albumin (BSA) for lipase immobilization, aimed at developing controlled enzymatic nano-drug delivery systems for targeted pancreatic cancer therapies. The primary objective is to enhance the kinetic stability and controlled release of lipase for hydrolyzing lipid membranes of cancer cells. The production of amyloid fibrils was optimized using Response Surface Methodology (RSM), with results from Congo Red spectroscopy (UV-Vis) and circular dichroism (CD) analyzed via Design-Expert ۷ software; the presence of nanostructures was confirmed by transmission electron microscopy (TEM). Immobilization was achieved through glutaraldehyde cross-linking, and kinetic parameters such as Km, Vmax, optimal temperature, optimal pH, and thermal stability of the immobilized lipase were compared with the free form. The results indicated that the highest amyloid yield was obtained after ۷۲.۶ hours of stirring at a BSA concentration of ۴.۳۵ mg/mL, pH ۴.۴۹, and ۸۰°C. The immobilized lipase exhibited superiority in Km and Vmax (decreased Km and increased Vmax), elevating the optimal temperature and pH to ۵۰-۶۰°C and ۷-۸, respectively, while demonstrating higher thermal stability at ۴۰°C; the nanofibril scaffolds (diameter <۱۰۰ nm) facilitated controlled release in the acidic microenvironment of pancreatic tumors. This biocompatible protein nanomatrix offers substantial potential for in vitro applications in therapeutic nanobiotechnology and significantly augments enzymatic properties.

کلیدواژه ها:

Amyloid ، immobilization ، Pancreatic cancer ، Nanotechnology