Sample Preparation Techniques for Mass Spectrometry in Proteomics Using Recently Developed Highly Selective Materials

Abstract

Efficient sample preparation in the micro and nano-litre range, prior to analysis of proteins and peptides of biological origin by liquid chromatography (LC), liquid chromatography coupled to mass spectrometry (LC-MS), micro-liquid chromatography (µ-LC) and matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry, is the key to success for different applications, such as biomarker discovery. Sample preparation in proteomics mainly comprises purification and preconcentration by solid-phase extraction (SPE) using different polymers (polystyrene, poly acrylate, cellulose, etc.) as a stationary phase following several cleaning-up procedures. These polymers can be derivatized with several functional groups, for example, C18, ion-exchanger or immobilized metal affinity chromatography (IMAC) groups. Based on these "traditional" techniques new methods enabling higher selectivity, sensitivity and speed of separation are required. Capillary coatings, for example, with latex particles has been used successfully. Derivatization of these latex particles, for example, with IMAC-functional groups enables a selective purification of phosphorylated or His-tagged proteins. Alternatively, polymeric disks possessing a thickness of approximately 2 mm can be used even in the case of multidimensional separations. In this review, we summarize currently available sample pretreatment techniques, provide an overview on the technical background and discuss the advantages of the individual methods using recently developed selective stationary phases such as silica based materials, polymers, etc.

Recent Developments in Solid-Phase Extraction for Near and Attenuated Total Reflection Infrared Spectroscopic Analysis

“…An evaluation of commercial available SPE materials for the selective analysis of monoterpenoids was reported by Uhlschmied et al [21]. For SPE of biomolecules the number of tailored stationary phases is even higher, e.g., for phosphopeptide and biomarker analysis, cellulose [22], poly(glycidyl-methacrylate/divinylbenzene) [23,24,25,26], carbon nanotubes [27], diamond layer [28,29], fullerenes [30,31,32,33], diamond-like-carbon [34,35], titanium and zirconium oxide [36], graphitic nanofibers [37], chitosan [38] and silica lanthanum oxide [39]. Nowadays the analysis after the enrichment step is mainly carried out by mass spectrometric (MS) techniques, including matrix assisted laser desorption ionization time of flight (MALDI-TOF) [40,41,42,43], electrospray ionization (ESI) and still high-performance liquid chromatography (HPLC) employing UV-absorbance, diode array detection and others [8].…”

Progress in capillary electrophoresis coupled to matrix‐assisted laser desorption/ionization – time of flight mass spectrometry

“…Therefore, several procedures have been described using SPE, membrane technology, disk technology, on-place preconcentration, and desalting. The advantages and disadvantages of the individual procedures have been exhaustingly discussed in a review provided by Huck et al [35].…”

Progress in Silica Chemistry - Determination of Physico-Chemical Parameters via Near-Infrared Diffuse Reflection Spectroscopy

“…(1) visualise the differences in morphology and application fields. Miniaturised separation systems such as micro liquid chromatography ( -LC) [4], capillary electrochromatography (CEC) [5] and material-enhanced laser desorption/ionisation time of flight mass spectrometry (MELDI-TOF/MS) [6], beside sample pre-treatment techniques [7] are emerging techniques, enabling high sensitivity, selectivity for e.g. biomarker discovery.…”