Optimization Of Bovine Serum Albumin Fibrillation By Congored Spectrophotometry For Using As A New Bio-Nanomaterial
Aim and Background: Amyloids are fibrilar protein structures produced from aggregation of proteins and
peptide together. In recent years, amyloid fibrils are being considered as new experimental nanomaterials and from this
perspective, increased levels of amyloid proteins can be desirable. In this study, the fibrillation of bovine serum albumin
(BSA) as a model protein is optimized.
Materials and Methods: Different concentrations of protein were subjected to different pH and temperature and amount of
amyloid fibrils were detected by spectrophotometric Congored binding assay. This dye can integrate to fibrous amyloid
structures and change the absorbance and λmax of fibrillated protein comparison to native form.
Results: 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.
Conclusion: 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.
Keywords— Bovine Serum Albumin, Amyloid, Spectrophotometry, Congored.