GEN-MKT-18-7897-A
Mar 28, 2018 | Blogs, Life Science Research, Lipidomics | 0 comments
Lipidomics research has progressed rapidly in the last decade. Twenty years ago, the determination of the lipid composition of a biological sample required slow and extensive multi-dimensional chromatographic separation and chemical derivatization strategies. Today, in-depth analysis can be performed quickly using high throughput analysis by mass spectrometry.
Advances in mass spectrometry have enabled in-depth lipidomic analyses with unparalleled qualitative and quantitative sensitivity. However, unambiguous identification and quantitation of lipid molecular species in total lipid extracts has proven to be difficult, primarily due to isobaric overlapping isobaric and isomeric species. There are greater than 100,000 lipid molecular species present in a typical biological lipid extract that occupy a narrow mass range (~400-1100 amu), making such overlap a significant problem.
SelexION Technology represents the next technological advance and is uniquely suited for lipidomic research. Using Differential Mobility Separation (DMS) as an upfront orthogonal separation enables the resolution of complex lipid mixtures, effectively addressing the problem of isobaric interferences among different lipid classes and sub-classes.
Key Features of SelexION Technology for Lipid Analysis
SelexION Technology is a planar differential mobility device that separates analytes based on differences in their chemical properties, prior to entering the instrument orifice, thus providing an orthogonal level of selectivity.
Read our technical paper to see how we effectively resolves multiple lipid classes from complex mixtures prior to analysis by mass spectrometry. By removing isobaric and isomeric interferences, lipid analysis by QTRAP systems using SelexION Technology enables more confident identification of lipid molecular species and provides for more accurate relative quantitation by MS/MS.
As an analytical strategy, middle-down mass spectrometry (MS) workflows characterize biotherapeutic proteins by analyzing large, digested protein fragments or defined subunits, rather than fully intact proteins (top-down) or digested peptides (bottom-up). A middle-down strategy combines the strengths of top-down and bottom-up approaches by delivering high sequence coverage and structural specificity while maintaining relatively simple sample preparation. In practice, middle-down analysis enables accurate mass measurement, rapid sequence confirmation, and localization of key post-translational modifications (PTMs) on protein subunits that are directly relevant to product quality.
In biopharmaceutical development, sequence variants (SV) are considered an inherent risk of producing complex proteins in living systems. Sequence variants are unintended changes to the amino acid sequence of a biotherapeutic and can be caused by errors in transcription or translation in the host cell, or cell culture and process conditions. Detailed analysis of SVs is important in process and product development to ensure the drug’s safety and efficacy. Even low‑level sequence variants can have significant implications for product quality, safety, and efficacy, making their accurate detection and characterization a critical requirement across development, process optimization, and regulatory submission.
CE‑SDS remains a cornerstone assay for characterizing fragmentation, aggregation, and product‑related impurities in therapeutic proteins. UV detection has been the long‑standing standard. However, it frequently struggles with baseline noise, limited sensitivity for minor fragments, and subjective integration.
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