Online SERS detection and characterization of eight biologically-active peptides separated by capillary zone electrophoresis

Abstract

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.