Automation of nanoflow liquid chromatography‐tandem mass spectrometry for proteome and peptide profiling analysis by using a monolithic analytical capillary column

Abstract

An automated nano-LC-MS/MS platform without trap column was established, which only used a 20 cm lauryl methacrylate-ethylene dimethacrylate (LMA-EDMA) monolithic capillary column to allow preconcentration and separation of peptides. The monolithic column had the advantages of good permeability and low backpressure resulting in higher flow rates for capillary columns. Tryptic digests of bovine albumin and yeast protein extract were tested using the monolithic column system. High proteomic coverage using this approach were demonstrated in this study. Furthermore, peptide samples extracted from mouse liver were separated by using the monolithic column system combined with size-exclusion chromatography prefractionation. This monolithic column system might be a promising alternative for the automated system previously using a trap column for routine proteome and peptide profiling analysis.

Monoliths in capillary electrochromatography and capillary liquid chromatography in conjunction with mass spectrometry

“…An automated cLC‐MS/MS platform without trap column was established in the group of Hanfa Zou . The monolithic poly(LMA‐EDMA) capillary column allowed simple preconcentration and separation of peptides.…”

Separation of intact proteins by using polyhedral oligomeric silsesquioxane based hybrid monolithic capillary columns

“…LMA-EDMA organic monolithic column was already applied for the peptides separation [26,27], and this organic monolith exhibited abundant mesopores and macropores, which may be suitable for the intact protein separation [26,28]. A total of 18 monolithic capillary columns with 100 m i.d.…”

Recent advances in monolithic columns for protein and peptide separation by capillary liquid chromatography

“…Various kinds of monolithic capillary columns have been synthesized for RPLC separation of proteins and peptides, such as poly(styrene-co-divinylbenzene) monoliths [53][54][55][56], poly(butyl methacrylate-co-ethylene dimethacrylate) monoliths [57], poly(lauryl methacrylateco-ethylene dimethacrylate) monoliths [58,59], monoliths derived from ring-opening metathesis polymerization [60], C 18 -modified silica monoliths [61][62][63][64][65][66], dimethyldodecylammonium-silica hybrid monoliths [44], POSS-based hybrid monoliths [49], and phenyl-silica hybrid monoliths [47]. The detailed applications are shown in Table 1.…”