Development of a high-throughput IMS–IMS–MS approach for analyzing mixtures of biomolecules

Abstract

A high-throughput approach for biomolecule analysis is demonstrated for a mixture of peptides from tryptic digest of four proteins as well as a tryptic digests of human plasma. In this method a chipbased electrospray autosampler coupled to a hybrid ion mobility (IMS) mass spectrometer (MS) is utilized to achieve rapid sample analysis. This high-throughput measurement is realized by exploiting the direct infusion capability of the chip based electrospray with its rapid sample manipulating capability as well as a high sensitive IMS-MS with a recently developed IMS-IMS separation technique that can be multiplexed to provide greater throughput. From replicate IMS-MS runs of known mixtures, the average uncertainty of peak intensities is determined to be ±7% (relative standard deviation), and a detection limit in the low attomole range is established. The method is illustrated by analyzing 124 human plasma protein samples in duplicate, a measurement that required 16.5 hours. Current limitations as well as implications of the high-throughput approach for complex biological sample analysis are discussed.

Utilising ion mobility-mass spectrometry to interrogate macromolecules: Factor H complement control protein modules 10–15 and 19–20 and the DNA-binding core domain of tumour suppressor p53

“…Early studies by Jarrold and Clemmer studied proteins such as ubiquitin [12] (∼8.5 kDa) and cytochrome c [13] (∼12.4 kDa) to characterise collision cross-sections under a variety of physical conditions such as temperature, pH and solvent conditions. More recent efforts have achieved greater sample throughput of biomolecule mixtures [14], resolving power [15] or sensitivity [16], allowing lower abundance species to be investigated. Instrument development has been integral to these achievements.…”

Delineating Diseases by IMS-MS Profiling of Serum N-linked Glycans

“…18,62 The specific conditions used in these studies are similar to those reported in our recent study of glycans. 30 General descriptions of theory and experiment that are valuable for understanding the concepts in this paper can be found elsewehere.…”

Overtone Mobility Spectrometry: Part 4. OMS-OMS Analyses of Complex Mixtures

“…The instrument used in this study is a modified version of existing IMS-MS instruments described previously [21, 43, 44]. The electrospray ionization (ESI) source and all the components that come after the OMS regions are unchanged.…”