The use of recombinant protein for therapeutic applications has increased significantly in the last three decades. The heterogeneity of these proteins, often caused by the complex biosynthesis pathways and the subsequent PTMs, poses a challenge for drug characterization to ensure its safety, quality, integrity, and efficacy. CE, with its simple instrumentation, superior separation efficiency, small sample consumption, and short analysis time, is a well-suited analytical tool for therapeutic protein characterization. Different separation modes, including CIEF, SDS-CGE, CZE, and CE-MS, provide complementary information of the proteins. The CE applications for recombinant therapeutic proteins from the year 2000 to June 2013 are reviewed and technical concerns are discussed in this article.
CE features superior separation efficiency, small solvent consumption, as well as the ability to analyze most biomolecules with an open tube fused-silica column. When coupled with MS, the separation power of CE is enhanced by adding another separation dimension based on mass-to-charge ratios. CE-MS reduces the dependence on CE separation so that faster analysis can be achieved. It also yields higher sensitivity as well as the capability for analyte identification and structural elucidation. The use of CE-MS for biomolecule analysis has increased significantly in the last 5 years. New methods are being developed for large molecules, while analyses of smaller molecules are moving toward the study in more complex tissues and other matrices. In this article, the applications of CE-ESI-MS for complex samples in 2007-2011 are reviewed. The applications are categorized according to the types of analytes studied, including the analysis for proteins and peptides, carbohydrates, and small biomolecules. Sample preparation methods, coatings for capillary inner wall, online processing strategies, and other aspects are also reviewed in each category.
Two novel supramolecular photoinitiators (supra-photoinitiators) based on the host−guest complexation of macrocycles (pillar[6]arene, P6; prism[5]arene, NP5) and diphenyl-iodonium salt (Iod) were fabricated. Under light irradiation, macrocycle P6 or NP5 could donate electrons to the guest molecule Iod, which generated highly active free radicals and cationic fragments to achieve efficient polymerization. Compared to a commercial activator, the electron transfer between macrocycles and Iod were under nondiffusion control, endowing a much higher photopolymerization rate and epoxy resin final conversion. In addition, the host−guest complexation of NP5 extended the initiating wavelength of Iod from ultrashort ultraviolet to near-ultraviolet, which could better match the environment-friendly LED light source. It is anticipated that a supra-photoinitiator may open a new route for designing novel photoinitiators with high performance.
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