Silica-based and organic monolithic capillary columns for LC: recent trends in proteomics.

The use of monolithic liquid chromatography (LC) columns for proteomics, covering the scientific literature from 2004 to the beginning of 2011, is reviewed. Attention is paid to recent developments in column technology and materials, focusing on silica-based and organic (polystyrene and methacrylate) monolithic capillary columns for proteomics. The applicability of these columns is illustrated by examples of the analysis of (complex) protein digests and proteins conveniently summarized in tables.

Comparison of monolithic and 1.8-μm RP-18 silica capillary columns using chromatographic data and mass spectrometric identification scores for proteins.

A goal in proteomics is the analysis of proteins by LC-MS. The proteins are enzymatically digested and the resulting peptides are chromatographically separated and introduced into a tandem MS. The obtained MS data are used for a search in sequence databases, providing identification scores for the proteins.

Development and characterization of cIEF-MALDI-TOF MS for protein analysis.

This paper describes the hyphenation of cIEF and MALDI-TOF MS via a fractionation or spotting device. After focusing in cIEF the compounds are hydrodynamically mobilized and deposited on a MALDI target plate using a sheath liquid interface, which provides the catholyte solution and the electrical ground. From previous experiments, sample conditions that resulted in a high resolution in cIEF and acceptable protein signal intensity in MS were selected [Silvertand et al.

Development of an on-line SPR-digestion-nanoLC-MS/MS system for the quantification and identification of interferon-gamma in plasma.

An automated, on-line system for protein quantification and identification, employing Surface Plasmon Resonance (SPR), enzymatic protein digestion, nanoLC and tandem-MS (MS/MS), has been developed. For the experiments recombinant human interferon-gamma (rhIFN-gamma) in buffer or diluted bovine plasma was used as a model protein. Upon injecting 90muL of a 1mugmL(-1) solution of rhIFN-gamma in diluted plasma at a flow rate of 10muLmin(-1), 320fmol of protein was reproducibly bound to the sensor surface.

Recent developments in capillary isoelectric focusing.

The developments in capillary isoelectric focusing (cIEF) over the period 2003-2007 are reviewed. With the focus on technological aspects, cIEF papers published in the fields of methodology, new techniques, detection, multidimensional systems, miniaturization and applications are summarized. The methodology section covers recent research in ampholytes composition, detergents and other additives, carrier ampholyte free cIEF, coatings and other capillary modifications.