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Optimizing plasma and serum sample preparation for deeper, high-throughput proteome profiling
Fast and efficient sample preparation workflows targeting low-abundance proteins in plasma, serum, CSF, and other high dynamic range samples for application in academic and biopharma research
The challenge
Accessing low-abundance proteins in plasma, serum, CSF and other high dynamic range samples for LC-MS-based proteomic analysis is very challenging due to the complexity and high dynamic range of proteins within these biological samples. High-abundance proteins such as e.g. albumin and immunoglobulins in plasma or serum make up a significant portion of the specimen and limit access to low-abundance proteins, which are more likely to represent relevant biomarkers in different applications.
The solution
PreOmics’ bead-based enrichment workflows allows to improve the high adynamic range challenge by increasing visibility of the underestimated fraction – low-abundance proteins – in plasma and serum samples. The all-in-one solution includes in addition to the enrichment step all core steps – enzymatic digestion and purification of resulting peptides by using the PreOmics® iST technology. The enrichment technologies enable processing of human samples and other mammalian species samples. With scalable throughput and exceptional reproducibility, the sample preparation takes only 5 hours from raw samples to pure peptides, and the analysis of large cohorts is possible with established lab automation systems suitable for magnetic bead processing. The technology offers an affordable solution that democratizes access to deep, unbiased plasma proteome analysis for diverse research needs.
Plasma, serum & CSF sample preparation simplified
ENRICH-iST is a flexible, fast and easy-to-use, robust all-in-one solution to prepare plasma, serum, CSF and other high dynamic range samples for proteomic analysis. It is based on the enrichment of low-abundance proteins onto paramagnetic beads and is compatible with human and other mammalian samples (e.g., mouse, rat, dog, pig, non-human-primate). The scalable throughput, exceptional reproducibility and a ultra-fast protocol from raw samples to pure peptides make ENRICH-iST a reliable tool for verifying and discovering biomarkers in large cohorts.
Results
Deeper proteome coverage compared to antibody-based depletion
According to the protein concentration from the Human Protein Atlas for plasma, ENRICH-iST offers superior analytical depth and proteome coverage.
Streamlined workflow for validation of known and novel biomarkers
ENRICH-iST identified 16 additional biomarkers in comparison to neat plasma.
Automatable, standardized workflow and higher reproducibility for reliable biological insights
ENRICH-iST shows a robust workflow with low technical variability: Protein CV of MaxLFQs < 20 % and equal quantitative performance between manual and automated sample preparation.
Redefining proteomics depth for plasma samples
P2-iST Plasma: Discover an innovative nanoparticle-based plasma proteomics solution designed for unprecedented depth and precision. Using P2-iST Plasma, ~6,000 proteins were identified in human EDTA plasma* samples with >90% data completeness across a pilot study of 12 cancer and healthy plasma samples. Built for scalability and efficiency, P2-iST enables high-throughput processing of up to 192 samples per day using standard automation platforms. To meet diverse research needs, flexible business models are available, ranging from research-use kits to dedicated CRO services.
P2-iST Plasma is currently available under Early Access, including comprehensive method transfer and implementation support for selected expert laboratories.
* Optimized for human EDTA plasma. Other anticoagulants or biofluids may result in different enrichment performance. Please consult with our technical experts.
P2-iST Plasma is currently available under Early Access, including comprehensive method transfer and implementation support for selected expert laboratories.
* Optimized for human EDTA plasma. Other anticoagulants or biofluids may result in different enrichment performance. Please consult with our technical experts.
Results
Figure 1. (Left) Streamlined plasma workflow combining PreOmics’ P2-iST Plasma sample preparation, Bruker's timsTOF HT mass spectrometer, and Biognosys Spectronaut® 20 software package. (Right) Compared to neat-prepared samples (iST-BCT), data from pooled human EDTA plasma* analyzed with a rapid 48SPD LC-method show up to an 8-fold increase in protein identifications with P2-iST.
What do our customers say?
"We tested several options for biomarker discovery on plasma samples, but many were long and tedious. With the novel ENRICH technology, we have a fast, efficient, and reliable enrichment workflow to get deeper into the proteome from human plasma and serum. We use the ENRICH-iST 96x kits and are delighted by the reproducibility and ease of the kit. We really enjoyed working with the PreOmics team, who was always there to support us. We would strongly recommend the kit to other plasma and serum users."
Dr. Catherine Nury
Manager Proteomics, Philip Morris International, Switzerland
“[…] ENRICHplus is changing the way we do plasma proteomics. On Bruker’s timsTOF Pro we could achieve on average 3,800 protein identifications per sample with a throughput of 60 samples per day while only using 20ul of plasma. We never before achieved such depth with a single run of plasma, now enabling us to detect tissue leakage proteins and other low abundant potential biomarkers for cancer and neurodegenerative diseases. […] ENRICHplus is a complete, efficient and game changing platform for deep plasma proteomics."
Roman Fischer
Associate Professor and Head of Discovery Proteomics Facility, University of Oxford
Key benefits of the enrichment technology
Enhanced proteome depth
Streamlined enrichment of low-abundance plasma proteins coupled with iST proteomic sample preparation technology for enhanced plasma proteome depth
Fast and reliable results
A fast, easy-to-use, and standardized protocol for high reproducibility and less hands-on time
Versatile technology
Species-independent and flexible technology compatible with plasma, serum, CSF, and other high dynamic range samples of different mammalian species
Automation friendly
Seamlessly integrated into standard automation platforms for processing up to 192 samples per day
Low input samples
Optimized technique to work with low-input starting material (20-100 μL) for large-scale proteomic studies
