Advances in microscale separations towards nanoproteomics applications

Abstract

Microscale separation (e.g., liquid chromatography or capillary electrophoresis) coupled with mass spectrometry (MS) has become the primary tool for advanced proteomics, an indispensable technology for gaining understanding of complex biological processes. In recent decades significant advances have been achieved in MS-based proteomics. However, the current proteomics platforms still face an analytical challenge in overall sensitivity towards nanoproteomics applications for starting materials of less than 1 µg total proteins (e.g., cellular heterogeneity in tissue pathologies). Herein, we review recent advances in microscale separation techniques and integrated sample processing strategies that improve the overall sensitivity and proteome coverage of the proteomics workflow, and their contributions towards nanoproteomics applications.

Modern separation techniques coupled to high performance mass spectrometry for glycolipid analysis

“…Because of such difficulties, though in the last few years important developments of CE ESI MS approaches, some even introducing chips and automation or combining the on‐line separation with top‐down fragmentation for protein characterization and metabolomics were reported, much less was achieved in innovations dedicated to glycoconjugate analysis. Still a number of original publications presenting excellent protocols and several reviews highlighting in details the advancements of the method for general glycomics are available .…”

New mass spectrometry technologies contributing towards comprehensive and high throughput omics analyses of single cells

“…However, consistent improvements in each of these areas has greatly increased the feasibility of in-depth proteome measurements for single cells and other small samples for which only picogram or low-nanogram amounts of protein starting material are available. 17,18 Below we highlight advances in sample handling, prior to MS analysis.…”

Portable capillary liquid chromatography for pharmaceutical and illicit drug analysis

“…In contrast to GC, the use of capillary‐scale columns has not dominated the LC field during its maturation. Rather, capillary LC has been primarily utilized in “omics” applications because of its enhanced sensitivity when interfaced to mass spectrometry (MS) as compared to analytical‐scale columns operated at much higher flow rates . The key advantages of capillary LC include (1) reduced flow rate, which makes the method greener compared to analytical‐scale LC, and (2) improved heat dissipation, which eliminates the effects of viscous friction when coupled with lower flow rates .…”