Abstract
In this study, a new method for the quantitative evaluation of proteins is developed using competitive resonance Raman spectroscopy. A chelation reaction between bathocuproine disulfonate (BCS) and Cu+ which is reduced by proteins in an alkaline environment, is utilized to create a BCS–Cu+ complex that has strong resonance Raman activity. As a result, the method presented here enables protein detection over a much wider concentration range than conventional methods. Furthermore, the resonance Raman-based method can provide an improved lower limit of detection (500 pg/mL) compared to that obtained by colorimetric and ultraviolet- (UV-) based methods. Additionally, protein-to-protein variation can be determined using a standard curve created from known concentrations of bovine serum albumin (BSA), and excellent protein recovery is observed. More importantly, the proposed method was employing to the real sample (fetal bovine serum [FBS]). Based on the calibration curve from this method, it is extremely accurate for the determination of total protein concentrations in FBS. Results of this study demonstrate that the proposed method provides a novel and highly sensitive protocol for reagent-based protein assays.
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