Recent advances in capillary separations for proteomics

Abstract

The sequencing of several organisms' genomes, including the human's one, has opened the way for the so-called postgenomic era, which is now routinely coined as "proteomics". The most basic task in proteomics remains the detection and identification of proteins from a biological sample, and the most traditional way to achieve this goal consists of protein separations performed by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). Still, the 2-D PAGE-mass spectrometry (MS) approach remains lacking in proteome coverage (for proteins having extreme isoelectric points or molecular masses as well as for membrane proteins), dynamic range, sensitivity, and throughput. Consequently, considerable efforts have been devoted to the development of non-gel-based proteome separation technologies in an effort to alleviate the shortcomings in 2-D PAGE while reserving the ability to resolve complex protein and peptide mixtures prior to MS analysis. This review focuses on the most recent advances in capillary-based separation techniques, including capillary liquid chromatography, capillary electrophoresis, and capillary electrokinetic chromatography, and combinations of multiples of these mechanisms, along with the coupling of these techniques to MS. Developments in capillary separations capable of providing extremely high resolving power and selective analyte enrichment are particularly highlighted for their roles within the broader context of a state-of-the-art integrated proteome effort. Miniaturized and integrated multidimensional peptide/protein separations using microfluidics are further summarized for their potential applications in high-throughput protein profiling toward biomarker discovery and clinical diagnosis.

Monolithic porous polymer stationary phases in polyimide chips for the fast high-performance liquid chromatography separation of proteins and peptides

“…Due to the complexity of proteomes and the small quantities of samples typically available, microsystems that afford high sensitivity, rapid throughput, and excellent reproducibility [2][3][4] designed to carry out the proteomic analysis. In this context, microfluidic systems integrating all parts of the separation device into a single chip are particularly attractive.…”

CE with a new electrochemiluminescent detection system for separation and detection of proteins labeled with tris(1,10‐phenanthroline) ruthenium(II)

“…CE possesses the advantages of high resolution, rapid separation and ease of automation. Therefore, the great potential of CE has been envisioned for the analysis of proteins and investigation of proteomes [1][2][3]. However, a major challenge in CE detection of protein is the poor concentration detection limit due to the short optical path length across the capillary and the small sample volumes injected.…”

A continuous wavelet transform algorithm for peak detection

“…[1][2][3][4]) due to the advantages of higher speed, higher sensitivity, smaller sample size, multiple sample analysis and on-column detection (e.g. [5]).…”