Negri Pierre, Sarver Scott A, Schiavone Nicole M, Dovichi Norman J, Schultz Zachary D
Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA.
Analyst. 2015 Mar 7;140(5):1516-22. doi: 10.1039/c4an01980f. Epub 2015 Jan 19.
There is a need for low cost, sensitive and chemical specific detectors for routine characterization of biomolecules. In this study, we utilize sheath-flow surface-enhanced Raman scattering (SERS) to analyze a mixture of eight biologically-active peptides separated by capillary zone electrophoresis (CZE). Analysis of the SERS electropherogram resulting from online detection resolves the characteristic Raman bands attributed to the amino acid constituents of each peptide, which enables identification. The detection limit by SERS was found to be 10(-8) M. Our results suggest that the structural information obtained from the detected vibrational modes provides complementary characterization to other chemically specific detectors like mass spectrometry and improved chemical identification over other commonly used optical-based post-chromatographic detection methods. In addition, the sheath-flow SERS detection results in band narrowing in the observed electropherogram that enables distinction of closely migrating species. The results presented here indicate that this platform can provide fast, robust, reproducible, and chemical specific detection to facilitate the characterization of peptides.
需要低成本、灵敏且具有化学特异性的探测器用于生物分子的常规表征。在本研究中，我们利用鞘流表面增强拉曼散射（SERS）来分析通过毛细管区带电泳（CZE）分离的八种生物活性肽的混合物。对在线检测得到的SERS电泳图的分析解析出了归因于每种肽氨基酸成分的特征拉曼带，从而实现了鉴定。发现SERS的检测限为10^(-8) M。我们的结果表明，从检测到的振动模式获得的结构信息为质谱等其他化学特异性探测器提供了互补的表征，并且比其他常用的基于光学的色谱后检测方法具有更好的化学鉴定能力。此外，鞘流SERS检测导致观察到的电泳图中谱带变窄，从而能够区分迁移相近的物种。此处呈现的结果表明，该平台可以提供快速、稳健、可重复且具有化学特异性的检测，以促进肽的表征。
