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Updated: Jul 9, 2026

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
Published on: November 15, 2017
Development of a comprehensive multidimensional liquid chromatography system with tandem mass spectrometry detection
Todd Kajdan1, Hernan Cortes, Krishna Kuppannan
1The Dow Chemical Company, Analytical Sciences, 1897 Building, Midland, MI 48667, USA.
Abstract:
A multidimensional comprehensive liquid-phase separation system (2DLC) coupled on-line to an electrospray-ionization time-of-flight (ESI-TOF) mass spectrometer (MS) was used to resolve structural alterations and/or post-translational modifications for detailed protein characterization. The system described in this work consists of cation-exchange chromatography in the first dimension and reversed-phase chromatography in the second dimension. A unique spiked gradient was employed in the first dimension to enhance recovery of peptides. This combination of separation followed by MS detection offered the advantages of unique selectivity and high efficiency of the separation methods combined with the mass specificity and sensitivity of MS. During the course of this study it was determined that altered or modified peptides were shown to be better resolved than during a one-dimensional separation. The 2DLC/ESI methodology allowed for a comprehensive evaluation of post-translational modifications and chemical reactions of recombinant proteins, providing a meaningful evaluation of product quality that was not possible with other current analytical approaches. In addition, the system can be used to provide sequence coverage of complex proteins.
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