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Coupling of capillary electrophoresis (CE) with mass spectrometry (MS) represents a powerful combination for performing rapid, efficient, and sensitive analysis of a variety of compounds. Here we describe a construction, operation, and application of a microfabricated liquid junction CE-MS interface. The interface is designed as a microfabricated unit with an integrated liquid junction and electrospray tip made from polyimide, which is positioned in a plastic connection block securing the separation CE capillary and attachable to the CE instrument. The application was demonstrated by CE-MS analysis of dextran oligomers labeled by (2-aminoethyl)trimethylammonium (AETMA) salt.

Surface-enhanced Raman spectroscopy (SERS) is an extremely powerful analytical tool, which not only yields information about the molecular structure of the analyte in the form of characteristic vibrational spectrum but also gives sensitivities approaching those in fluorescence spectroscopy. The SERS measurement on the microfluidic platform provides possibility to manufacture the device with design perfectly fulfilling the needs of the application with minimal sample consumption. This review aims at describing basic strategies for SERS measurement in microfluidic devices published in the last decade and covers current trends in microfluidics with SERS detection in the field of bioanalysis and approaches toward on-line coupling of liquid-based separation techniques with SERS detection.

The direction of the effective electrophoretic mobility of small organic cations in micellar electrokinetic chromatography using sodium dodecyl sulphate in a low-pH electrolyte can be reversed in the presence of organic solvent. This effective electrophoretic mobility change is presented here as a new dimension for on-line sample preconcentration of cations in capillary zone electrophoresis (CZE) using a background solution (BGS) modified by an organic solvent. The sample is prepared in a micellar solution without organic solvent. The focusing effect relies on the reversal in the effective electrophoretic mobility at the boundary zone between the micellar matrix and the BGS modified with organic solvent. This on-line sample preconcentration technique, called micelle to solvent stacking (MSS) afforded more than an order of magnitude improvement in concentration sensitivity compared to typical CZE-UV or CZE-electrospray ionization (ESI) MS analysis. The calculated limit of detection (S/N=3) for pindolol and metoprolol analysed by MSS-CZE-ESI-MS was found to be 0.03 and 0.01 microg/mL, respectively.

The use of high quality fused silica capillary nanospray tips is critical for obtaining reliable and reproducible electrospray/MS data; however, reproducible laboratory preparation of such tips is a challenging task. In this work, we report on the design and construction of low-cost grinding device assembled from 3D printed and commercially easily available components. Detailed description and characterization of the grinding device is complemented by freely accessible files in stl and skp format allowing easy laboratory replication of the device. The process of sharpening is aimed at achieving maximal symmetricity, surface smoothness and repeatability of the conus shape. Moreover, the presented grinding device brings possibility to fabricate the nanospray tips of desired dimensions regardless of the commercial availability. On several samples of biological nature (reserpine, rabbit plasma, and the mixture of three aminoacids), performance of fabricated tips is shown on CE coupled to MS analysis. The special interest is paid to the effect of tip sharpness.

We report a construction of a self-aligning subatmospheric hybrid liquid junction electrospray interface for CE eliminating the need for manual adjustment by guiding the capillaries in a microfabricated liquid junction glass chip at a defined angle. Both the ESI and separation capillaries are inserted into the microfabricated part until their ends touch. The distance between the capillary openings is defined by the angle between the capillaries. The microfabricated part contains channels for placement of the capillaries and connection of the external electrode reservoirs. It was fabricated using standard photolithographic/wet chemical etching techniques followed by thermal bonding. The liquid junction is connected to a subatmospheric electrospray chamber inducing the flow inside the ESI needle and helping the ion transport via aerodynamic focusing.

There are a considerable number of biopharmaceuticals that have been approved for clinical use in the past decade. Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. The linked carbohydrate moieties affect not only their physicochemical properties and thermal stability but also crucial features like receptor-binding activity, circulating half-life, as well as immunogenicity. The structural diversity of these attached glycans can be manifested in altered monosaccharide composition and linkages/positions among the monosaccharide building blocks. In addition, as more and more biosimilar products hit the market, understanding the effects of their glycosylation modification has become a recent target in efficacy and safety issues. To ensure consistent quality of these products, glycosylation profiles have to be monitored and controlled in all steps of the manufacturing process, i.e., from clone selection to lot release. In this paper, we describe some of the recently introduced and commonly used sample preparation techniques for capillary electrophoresis (CE)-based profiling and structural elucidation of N-glycans. The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans.

In this work we have developed a hydrophilic poly(hydroxyethyl methacrylate-co-poly(ethylene glycol) diacrylate) cryogel placed in the centrifugal filter device. The composition of the polymerization mixture as well as the polymerization conditions were optimized in order to prepare a material with bimodal pore size distribution with 20-50 μm flow through macropores and submicrometer pores in the polymer walls. The optimized, mechanically stable, highly porous, material was used for spin column lectin chromatography. The surface of the monolithic scaffold was activated by epichlorohydrin and used for immobilization of concanavalin A to provide the affinity supports for selective isolation of glycoproteins containing high mannose glycan structures. The performance of the developed lectin modified cryogels was evaluated by analyses of glycoprotein mixtures. The efficiency and selectivity of the affinity supports were confirmed by MALDI-MS analysis.

A microfabricated pneumatic electrospray nebulizer has been developed and evaluated using computer simulations and experimental measurements of the MS signals. The microdevice under development is designed for electrospray MS interfacing without the need to fabricate an electrospray needle and can be used as a disposable or an integral part of a reusable system. The design of the chip layout was supported by computational fluid dynamics simulations. The tested microdevices were fabricated in glass using conventional photolithography, followed by wet chemical etching and thermal bonding. The performance of the microfabricated nebulizer was evaluated by means of TOF-MS with a peptide mixture. It was demonstrated that the nebulizer, operating at supersonic speed of the nebulizing gas, produced very stable nanospray (900 nL/min) as documented by less than 0.1% (SE) fluctuation in total mass spectrometric signal intensity.

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Capillary electrophoresis hyphenated with electrospray mass spectrometry (CE-ESI-MS) has emerged in the past decade as one of the most powerful bioanalytical techniques. As the sensitivity and efficiency of new CE-ESI-MS interface designs are continuously improving, numerical modeling can play important role during their development. In this review, different aspects of computer modeling and simulation of CE-ESI-MS interfaces are comprehensively discussed. Relevant essentials of hydrodynamics as well as state-of-the-art modeling techniques are critically evaluated. Sheath liquid-, sheathless-, and liquid-junction interfaces are reviewed from the viewpoint of multidisciplinary numerical modeling along with details of single and multiphase models together with electric field mediated flows, electrohydrodynamics, and free fluid-surface methods. Practical examples are given to help non-specialists to understand the basic principles and applications. Finally, alternative approaches like air amplifiers are also included. © 2014 Wiley Periodicals, Inc. Mass Spec Rev 34: 558-569, 2015.

Scaffolds modified with nanostructures are recently finding use in a broad range of applications spanning from chromatographic separations to tissue engineering. This continuation of the review series on design and applications of monolithic materials covers some of the less common monoliths including use of nanostructures in preparation, modifications, and applications.

Body fluids and body tissues have a myriad of peptides and proteins that, very often, the traditional methodologies of proteomics, such as conventional gel electrophoresis or mass spectrometry, are unable to characterize. We describe two protocols to characterize high molecular weight peptides (3 kDa) and intact proteins involving on-line trypsin digestion, separation of the digests by nano-HPLC, and analysis by mass spectrometry using two different ionization sources (matrix-assisted laser desorption and electrospray ionization). These protocols have the advantage of promoting protein denaturation in an aqueous-organic solvent, which reduces the derivatization of the sample and facilitates an in-depth analysis for detection and identification of proteins. Additional advantages include the following: (1) integration of these protocols into standard proteomic workflows after the preprocessing of samples and separation; (2) use of high-resolution monolithic columns; and (3) the ability to acquire information from minimal amounts of sample.

Advanced metal deposition and microfabrication techniques enable preparation of metal surfaces with high precision and excellent control over their size and shape with subnanometer resolution. Thin metal films of different types and functions can be found in many analytical instruments. Surfaces with high optical quality serve as mirrors, beam splitters, antireflective coatings etc. Smooth metal coating is crucial in electron microscopy. Unique properties of the thin metal films are widely used in optical systems, as tools for sample manipulation but also for chemical sensing and detection. While some of the applications are widespread and belong to the basic curriculum in analytical chemistry, the newer or less common uses of thin metal films are well known only to the experts in the field. The purpose of this critical review is to highlight the role of thin metal films in bioanalysis and summarize some of their main applications in current bioanalytical instrumentation.

In the present work, an exhaustive review of the main developments and applications of CEMS for peptide analysis is given. This review includes the use of different CE separation modes, MS analyzers, capillary coatings, preconcentration techniques, on-chip applications as well as other different multidimensional strategies for peptide analysis. Key applications are critically discussed and relevant works published from January 2000 to May 2007 are summarized including information concerning the type of sample, CE-MS parameters as well as some figures of merit of the different CE-MS procedures developed for peptide analysis and peptidomics., This study was supported by ProjectsAGL2005-05320-C02-01 Ciencia) and S-505/AGR-0153 (ALIBIRD, Comunidad de Madrid). M. H. would like to thank the Spanish Education and Science Ministry (MEC) for a post-doctoral fellowship.

Thin metal layers are widely utilized in a number of modern applications in electronics, optics, chemistry and many other applications. At present the thin layers of metals serve in applications spanning from simple electrodes to sur- face plasmon resonance (SPR) or giant magnetoresistance (GMR) based sensors (1, 2). Thin films allow monitoring redox processes in the vicinity of electrodes, adsorption/desorption equilibria of ions, organic compounds, gases, and more re- cently also interactions of large organic macromolecules such as proteins or DNA. In a previous review we have covered applications of thin films in biosensing regardless of the measured physical change (3). Measurement of basic electric properties (current, voltage, resistance) is simple and precise with large dynamic range. Since the related equipment is simple and inexpensive; such sensors might be very attractive for many applications in chemistry, environmental sciences or medicine. In this work, we shall discuss the potential of metal layers for future applications in resistance based chemi- cal sensing.