Analysis of proteins and peptides by electromigration methods in microchips

Abstract

This review presents the developments and applications of microchip electromigration methods in the separation and analysis of peptides and proteins in the period 2011-mid-2016. The developments in sample preparation and preconcentration, microchannel material, and surface treatment are described. Separations by various microchip electromigration methods (zone electrophoresis in free and sieving media, affinity electrophoresis, isotachophoresis, isoelectric focusing, electrokinetic chromatography, and electrochromatography) are demonstrated. Advances in detection methods are reported and novel applications in the areas of proteomics and peptidomics, quality control of peptide and protein pharmaceuticals, analysis of proteins and peptides in biomatrices, and determination of physicochemical parameters are shown.

Industrial bioprocessing perspectives on managing therapeutic protein charge variant profiles

“…Thus far, much of the development remains in the academic arena, and challenges preventing commercial usage include cost, lack of robustness, and difficulty in operation. However, tremendous progress has been made in the past few years that may accelerate eventual adoption by the biopharmaceutical industry (Štěpánová & Kašička, ).…”

“…Thus far, much of the development remains in the academic arena, and challenges preventing commercial usage include cost, lack of robustness, and difficulty in operation. However, tremendous progress has been made in the past few years that may accelerate eventual adoption by the biopharmaceutical industry (Štěpánová & Kašička, 2017 Zhou, 2012), and is selected based on the protein loading and binding needed to achieve flow rates at 100-500 cm/hr and a load capacity as high as 100 mg/ml . Despite these limitations however, PD groups may still be able to affect charge variant separation by resin choice.…”

Recent progress of sample stacking in capillary electrophoresis (2016–2018)

“…General reviews were aimed at enrichment techniques and enhancement of sensitivity in CE , their application in chiral CE , in CE‐MS and in the analysis of alkaloids . Electrophoretic stacking was also included in specialized reviews on CE analyses of peptides and proteins , pharmaceuticals , pesticides , antibiotics , food , and biologically important thiols . The use stacking by pH junction in CE‐ESI‐MS for proteomic analyses was reviewed in .…”

Determination of protein charge variants with (imaged) capillary isoelectric focusing and capillary zone electrophoresis

“…Through the whole‐capillary imaging, advantageously no mobilization is necessary. Isoelectric focusing can also take place on a chip with microfluidic technology . This leads to a high‐throughput and even lower sample concentrations.…”