Do the complementarities of electrokinetic and chromatographic procedures represent the “Swiss knife” in proteomic investigation? An overview of the literature in the past decade

Abstract

This report reviews the literature of the past decade dealing with the combination of electrokinetic and chromatographic strategies in the proteomic field. Aim of this article is to highlight how the application of complementary techniques may contribute to substantially improve protein identification. Several studies here considered demonstrate that exploring the combination of these approaches can be a strategy to enrich the extent of proteomic information achieved from a sample. The coupling of "top-down" and "bottom-up" proteomics may result in the generation of a hybrid analytical tool, very efficient not only for large-scale profiling of complex proteomes but also for studying specific subproteomes. The range of applications described, while evidencing a continuous boost in the imagination of researchers for developing new combinations of methods for protein separation, also underlines the adaptability of these techniques to a wide variety of samples. This report points out the general usefulness of combining different procedures for proteomic analysis, an approach that allows researchers to go deeper in the proteome of samples under investigation.

Sodium dodecyl sulfate free gel electrophoresis/electroelution sorting for peptide fractionation

“…Technological advancements in instrumentation, coupling to mass spectrometer and improvements in separation performance have accompanied this development . Even though nanoLC–MS is still the most commonly used analysis platform for advanced proteomics , several studies have demonstrated that combination of nanoLC–MS and CE–MS methods increase the number of identified proteins as they are orthogonal and provide complementary results .…”

Sample preparation and fractionation techniques for intact proteins for mass spectrometric analysis

“…CE‐MS has matured since the early work from McLafferty and coworkers [155] into a robust analytical tool capable of deep examination of complex proteomes and protein biomarkers in biological matrices [156,157]. Moreover, when it is applied together with LC‐MS, it can considerably increase protein coverage, as these techniques are orthogonal and provide complementary data [158]. CE can be coupled off‐line to MALDI‐MS and, as MALDI‐MS is more tolerant to sample constituents like buffers and salts, nonvolatile background electrolytes can be used for CE‐MALDI‐MS [159].…”

<p>Proteomic Applications in Antimicrobial Resistance and Clinical Microbiology Studies</p>

“…29 Another disadvantage of these approaches is their poor performance in detecting different types of post-translational modifications of a single protein that causes crosstalk among signal pathways. 30 While one of the disadvantages of membrane proteomics based on the gel-free approach is the solubilization of membranous proteins, which is because of different optimum condition, 31 the volume of data available for membrane protein repertoire is growing. 32 Several bacterial studies including Mycobacterium tuberculosis, 33 Scheffersomyces stipitis, 34 and Staphylococcus aureus 35 have used a gelfree technique, which further indicates the potential of this method by the identification of a far larger number of proteins.…”