Highly Sensitive Protein Concentration Assay over a Wide Range via Surface-Enhanced Raman Scattering of Coomassie Brilliant Blue
Highly Sensitive Protein Concentration Assay over a Wide Range via Surface-Enhanced Raman Scattering of Coomassie Brilliant Blue
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DOI：
10.1021/ac100596u
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发表时间：
2010-06-01
影响因子：
7.4
通讯作者：
Ozaki, Yukihiro
中科院分区：
文献类型：
--
作者：
Han, Xiao Xia;Xie, Yunfei;Ozaki, Yukihiro
在 Bradford 蛋白质测定中，由于考马斯亮蓝 G-250 (CBBG) 与蛋白质的结合，通过 595 nm 处的吸光度确定蛋白质浓度。在蛋白质-CBBG液体混合物中，表面增强拉曼散射（SERS）对吸附在银表面上的未结合CBBG分子的量敏感，并且结合的CBBG量与目标蛋白质浓度直接相关。因此，基于未结合的 CBBG 的 SERS 和硅内标，开发了一种检测溶液中总蛋白浓度的新方法。与传统的 Bradford 蛋白质测定相比，所提出的蛋白质测定法有两个明显的优点，即其更宽的线性浓度范围（10(-5)-10(-9) g/mL）和 200 倍的检测下限（1 ng/mL），这表明其在快速、高灵敏度测定高丰度和低丰度蛋白质浓度方面具有巨大潜力。
In the Bradford protein assay, protein concentrations are determined by the absorbance at 595 nm due to the binding of Coomassie brilliant blue G-250 (CBBG) to proteins. In a protein-CBBG liquid mixture, surface-enhanced Raman scattering (SERS) is sensitive to the amount of unbound CBBG molecules adsorbed on silver surfaces, and the bound CBBG amount is directly related to the target protein concentration. Accordingly, a novel method for detecting total protein concentration in a solution has been developed based on SERS of unbound CBBG with an internal standard of silicon. Two obvious advantages of the proposed protein assay over conventional Bradford protein assay are its much wider linear concentration range (10(-5)-10(-9) g/mL) and 200 times lower limit of detection (1 ng/mL), which demonstrates its great potential in rapid, highly sensitive concentration determination of high and low-abundance proteins.