Fully Automated Chip-Based Mass Spectrometry for Complex Carbohydrate System Analysis

Abstract

Carbohydrates represent a major class of biopolymers, which occur in nature either as oligosaccharides or glycoconjugates, in which the sugar moiety is linked to proteins or lipids. The significance of mass spectrometry for highly sensitive analysis of complex carbohydrates increased after the introduction of the electrospray ionization and matrix assisted laser desorption/ionization methods and the possibility of tandem MS for sequencing of single molecular species in complex mixtures. Rapid and sensitive characterization of carbohydrates in biological systems by automated nanoscale liquid delivery and chip-based electrospray interface techniques have not been developed so far. In this contribution, the implementation and optimization of a fully automated chip-based nanoelectrospray assembly (NanoMate system), operating in the negative ion mode, in combination with QTOF-tandem MS for mapping/sequencing and computer-assisted structure assignment for carbohydrate components in complex mixtures is presented.

0,2An cross-ring cleavage as a general diagnostic tool for glycan assignment in glycoconjugate mixtures

“…The possibility to obtain distinct fragmentation patterns under controlled collision energy and gas conditions along with the ability of automatic switching between MS and MS/MS mode by electrospray quadrupole time-of-flight Q-TOF instrument, represents a powerful option for high-throughput analysis of complex carbohydrate mixtures. This technique can be another option for such rapid investigation in combination with fully automated chip-ESI-QTOF-MS for high-performance glycoscreening and sequencing, developed by our group [14].The electrospray ionization followed by formation of fragment ions by collision-induced dissociation allows sensitive mapping and sequencing of oligosaccharides for identification. In general, carbohydrates undergo two different types of fragmentation: those of glycosidic cleavages and those of cross-ring fragmentation.…”

“…The possibility to obtain distinct fragmentation patterns under controlled collision energy and gas conditions along with the ability of automatic switching between MS and MS/MS mode by electrospray quadrupole time-of-flight Q-TOF instrument, represents a powerful option for high-throughput analysis of complex carbohydrate mixtures. This technique can be another option for such rapid investigation in combination with fully automated chip-ESI-QTOF-MS for high-performance glycoscreening and sequencing, developed by our group [14].…”

Chip electrospray mass spectrometry for carbohydrate analysis

“…For carbohydrate analysis, the NanoMate robot was first mounted onto the QTOF and IT mass spectrometers [47][48][49] via special brackets, allowing adjustment of the robot position with respect to the sampling cone. Recently, the first coupling of the NanoMate robot to the FTICR-MS at 9.4 T and sustained off-resonance CID (SORI CID) MS/ MS was reported [50].…”

“…A urine fraction containing sialylated carbohydrates from a patient presenting a clinical picture associated congenital disorders of glycosylation (CDG) was submitted to glycoscreening by fully automated chip-based (2)nanoESI/ QTOF-MS [47]. The sample with average concentration of 5 pmol/mL in methanol was loaded onto the 96-well plate of the NanoMate robot.…”

“…15). For preliminary analysis and assignment of glycoforms, a computer algorithm for calculation of glycoconjugate composition was developed [47,56] thus providing the first two-stages platform for completely automatic and highthroughput analysis: (1) robotized sample delivery and infusion into MS followed by (2) automatic identification of components using software-assisted assignment. The experimental data for the input were the m/z values and the respective charge states provided by chip MS and MS/MS experiments.…”