Top-Down Proteomics: Ready for Prime Time?

Abstract

TOC

Identification and Quantification of Proteoforms by Mass Spectrometry

“…A wide variety of fractionation and separation techniques have been combined to reduce the complexity of samples delivered to the mass spectrometer . Here, we highlight such techniques (summarized in Table ), including size‐based separations, high‐ and low‐pH RPLC, and CZE.…”

“…Additionally, other chromatography modes, such as hydrophobic interaction chromatography (HIC) and ion exchange chromatography (IEX), have been reported (see Chen et al.) . Another related and emerging separation in top‐down proteomics is ion mobility spectrometry …”

“…Top‐down proteomics directly analyzes intact proteins . The relationship between amino acid sequence and PTMs is preserved and proteoforms can be characterized, providing a proteoform‐specific understanding of biological phenomena.…”

“…The relationship between amino acid sequence and PTMs is preserved and proteoforms can be characterized, providing a proteoform‐specific understanding of biological phenomena. Recent reviews have covered sample preparation and mass spectrometry instrumentation techniques for top‐down proteomic analyses . Top‐down proteomic experiments are subject to many analytical challenges owing to the low abundance of many proteoforms and the low signal‐to‐noise (S/N) ratios inherent to mass measurement of large molecules .…”

FLASHDeconv: Ultrafast, high-quality feature deconvolution for top-down proteomics

“…Top-down (TD) proteomics has gained a lot of momentum for in-depth protein characterization and protein species analytics [1][2][3][4][5][6][7] . In contrast to bottom-up (BU) proteomics, where proteins are enzymatically digested and actually peptides are analyzed, the TD approach allows for the analysis of intact proteoforms (protein species arising from the same gene product via splice variants, genomic variation, post-translational modifications, degradation, etc.)…”