Issue 24, 2012

Quantitative evaluation of proteins with bicinchoninic acid (BCA): resonance Raman and surface-enhanced resonance Raman scattering-based methods

Abstract

A rapid and highly sensitive bicinchoninic acid (BCA) reagent-based protein quantitation tool was developed using competitive resonance Raman (RR) and surface-enhanced resonance Raman scattering (SERRS) methods. A chelation reaction between BCA and Cu+, which is reduced by protein in an alkaline environment, is exploited to create a BCA–Cu+ complex that has strong RR and SERRS activities. Using these methods, protein concentrations in solutions can be quantitatively measured at concentrations as low as 50 μg mL−1 and 10 pg mL−1. There are many advantages of using RR and SERRS-based assays. These assays exhibit a much wider linear concentration range and provide an additional one (RR method) to four (SERRS method) orders of magnitude increase in detection limits relative to UV-based methods. Protein-to-protein variation is determined using a reference to a standard curve at concentrations of BSA that exhibits excellent recoveries. These novel methods are extremely accurate in detecting total protein concentrations in solution. This improvement in protein detection sensitivity could yield advances in the biological sciences and medical diagnostic field and extend the applications of reagent-based protein assay techniques.

Graphical abstract: Quantitative evaluation of proteins with bicinchoninic acid (BCA): resonance Raman and surface-enhanced resonance Raman scattering-based methods

Supplementary files

Article information

Article type
Paper
Submitted
30 May 2012
Accepted
07 Oct 2012
First published
09 Oct 2012

Analyst, 2012,137, 5834-5838

Quantitative evaluation of proteins with bicinchoninic acid (BCA): resonance Raman and surface-enhanced resonance Raman scattering-based methods

L. Chen, Z. Yu, Y. Lee, X. Wang, B. Zhao and Y. M. Jung, Analyst, 2012, 137, 5834 DOI: 10.1039/C2AN35715A

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