Label-Free CE-MS Unveils Native N-Glycome in Single Cells and ng-Level Blood Samples
Summary
Researchers have developed a method that's akin to a miniature lab, remarkably compact yet hugely powerful, able to delve into single cells to see what's happening. This isn't just any routine procedure; it's a novel methodology that seamlessly integrates in-capillary sample processing with something called capillary electrophoresis-mass spectrometry. What it means in plain English is that they can now analyze these complex molecules in individual cells with incredible precision. In their research, they looked at HeLa and U87 cells - yes, those are types of cells used frequently in scientific studies - and found up to 100 different N-glycans in each cell. It's like uncovering a whole new layer of cellular identity.
Fig. 1 CE-MS-based experimental workflow for N-glycan profiling of single mammalian cells and blood-derived isolates.1
Research Criteria
They've come up with this new technique using capillary electrophoresis-mass spectrometry, or CE-MS for short, that's kind of a game-changer for studying tiny biological samples. It's specifically designed to profile N-glycans, which are crucial molecules involved in numerous biological processes like cell interaction and immune responses.
Sample Type
The researchers used HeLa and U87 cells. They were looking at how these cells change on a very detailed level after being treated with something called lipopolysaccharide, or LPS for short.
Result - Blood's N-Glycome Profiling
This new method allows for the detailed profiling of N-glycans from just tiny amounts of blood-derived samples. This technology can unravel the complex glycan structures found in human serum immunoglobulins like IgM and IgG, total plasma, and even plasma-derived extracellular vesicles (EVs). It's like having a super-microscope that lets us see the molecular intricacies that were previously a blur.
Fig. 2 CE-MS-based N-glycan profiling of human blood-derived isolates.1
Result - Single-Cell N-Glycome Changes Triggered by LPS Activation
Scientists have created a method known as CE MS to study the N glycomes of cells. They conducted this analysis on two types of cells; HeLa cells, commonly used in studies and U87 cells a type of brain cancer cell. The U87 cells displayed compositions compared to HeLa cells showing higher levels of fucosylation and sialylation. Additionally, the researchers investigated how these cells responded to stress induced by, lipopolysaccharide (LPS) a molecule that mimics infection. Each cell type exhibited changes in their profiles in response to the stressor. This detailed exploration could revolutionize our understanding of diseases and aid, in drug development.
Fig. 3 Differential qualitative and quantitative N-glycan profiling of single HeLa and single U87 mammalian cells.1
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Reference
- Marie, Anne-Lise, et al. "Native N-glycome profiling of single cells and ng-level blood isolates using label-free capillary electrophoresis-mass spectrometry." Nature communications 12.1 (2024): 3847.
