Optimization of bovine serum albumin fibrillation by congored spectrophotometric method for using as nanobiomaterial

Aim and Amyloids are fibrilar protein structures produced from aggregation of proteins and peptide together. In recent years amyloid fibrils are being noticed as new experimental nanomaterials and from this view the induction of proteins to amyloids could be beneficial. In this study, the fibrillation of bovine serum albumin (BSA) as a model protein is optimized.
Different pHs of 1, 2, 3, 4.7 and 7.4 of Bovine Serum Albumin were prepared in Mixed Citrate–Phosphate buffer and agitated for 0, 24, 48 and 72 hour in 100 RPM in various temperatures (40–80 ºC) for preparing protein fibrils. Then the amount of amyloid fibrils was detected by spectrophotometric congored binding assay.
In this research, effect of four variables include temperature, pH, time of incubation and protein concentration was investigated on fibrillogenesis and results were confirmed by congored spectrophotometric method as λmax and absorbance in λmax (Aλmax) accompanying with transmission electron microscopy. The optimum condition for fibrillogenesis was specified in 5 mg.ml-1 of protein and buffer pH of 3 after 72 hour incubation in 70ºC.
Simple congored spectrophotometric method could be used as primary test for evaluating protein nanobiofibrils and absorbance in λmax is introduced as a valid indicator in this way.