ReviewJournal of proteomics2024
Optimal conditions for carrying out trypsin digestions on complex proteomes: From bulk samples to single cells.
Review in Journal of proteomics, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed.
- A Uniquome-Based Method for Proteins Identification by Mass Spectrometry.Methods in molecular biology (Clifton, N.J.) · 2027Article
- Article
- Comparison of In-Gel and SP3 Based Sample Preparation Protocols for LC-MS/MS Based Proteomics.Proteomics · 2026Article
- Trypsin exhibits exopeptidase-like activity toward N-terminal arginine that biases proteomic analyses.bioRxiv : the preprint server for biology · 2026Article
- A Native Nepenthesin Reactor for Improved Proteolytic Digestion of Intrinsically Disordered Proteins in Proteomics Workflows.Chembiochem : a European journal of chemical biology · 2026Article
- Recent Developments and Applications of Capillary and Microchip Electrophoresis in Proteomics and Peptidomics (2023-2025).Journal of separation science · 2026Review
- Comparative evaluation of immunoaffinity and antibody-free isotope dilution mass spectrometry approaches for plasma ProGRP quantification.Analytical and bioanalytical chemistry · 2026Article
- Characterization of Lysine Methylation During Neuronal Differentiation of LUHMES cells.bioRxiv : the preprint server for biology · 2026Article
- Recent Developments in Capillary and Microchip Electroseparations of Peptides (2023-mid 2025).Electrophoresis · 2026Review
- Analysis of proteins and peptides to investigate the molecular signatures of a conventional antidiabetic herb, Boerhavia procumbens Banks ex Roxb. (Nyctaginaceae).Scientific reports · 2025Article
- Glial cells are involved in day-night protein dysregulation in the hippocampus of a mouse model of Alzheimer's disease.Scientific reports · 2025Article
- Bovine Serum Albumin-Trypsin Sponges for Enhanced Enzymatic Stability and Protein Digestion Efficiency.ACS applied bio materials · 2025Article
- CySP3-96 Enables Scalable, Streamlined, and Low-Cost Sample Preparation for Cysteine Chemoproteomic Applications.Molecular & cellular proteomics : MCP · 2025Article
- Development of an Adaptive, Economical, and Easy-to-Use SP3-TMT Automated Sample Preparation Workflow for Quantitative Proteomics.Journal of proteome research · 2025Article
- Comparison of Three DNA Isolation Methods and Two Sequencing Techniques for the Study of the Human Microbiota.Life (Basel, Switzerland) · 2025Article
- CD39 Contributes to the Ability of Cell Invasion in Heterogeneity of Colorectal Cancer.Journal of cellular and molecular medicine · 2025Article
- Proteomic Analysis of the Characteristic Flavor Components inFoods (Basel, Switzerland) · 2024Article
- Proteomics-The State of the Field: The Definition and Analysis of Proteomes Should Be Based in Reality, Not Convenience.Proteomes · 2024Article
Corrections and comments
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Authors and funding
5 authors.
Funding
Abstract
To identify proteins by the bottom-up mass spectrometry workflow, enzymatic digestion is essential to break down proteins into smaller peptides amenable to both chromatographic separation and mass spectrometric analysis. Trypsin is the most extensively used protease due to its high cleavage specificity and generation of peptides with desirable positively charged N- and C-terminal amino acid residues that are amenable to reverse phase HPLC separation and MS/MS analyses. However, trypsin can yield variable digestion profiles and its protein cleavage activity is interdependent on trypsin source and quality, digestion time and temperature, pH, denaturant, trypsin and substrate concentrations, composition/complexity of the sample matrix, and other factors. There is therefore a need for a more standardized, general-purpose trypsin digestion protocol. Based on a review of the literature we delineate optimal conditions for carrying out trypsin digestions of complex proteomes from bulk samples to limiting amounts of protein extracts. Furthermore, we highlight recent developments and technological advances used in digestion protocols to quantify complex proteomes from single cells. SIGNIFICANCE: Currently, bottom-up MS-based proteomics is the method of choice for global proteome analysis. Since trypsin is the most utilized protease in bottom-up MS proteomics, delineating optimal conditions for carrying out trypsin digestions of complex proteomes in samples ranging from tissues to single cells should positively impact a broad range of biomedical research.
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Registered trials
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