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New Kits and Novel Workflow for Plasma and Serum Proteomics by LC-MS

ENRICH-iST enables unbiased enrichment of lower abundant plasma and serum proteins

the ENRICH-iST kit on a black background

The technology enables biologically unbiased enrichment of lower abundant proteins and supports liquid handling automation platforms.

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PreOmics

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HOUSTON, TX — At ASMS 2023, PreOmics GmbH announces the launch of its new ENRICH-iST™ kits, which enable biologically unbiased enrichment of lower abundant plasma and serum proteins onto non-functionalized paramagnetic particles, enhancing typical depths of plasma proteomics analysis by 50 percent to more than 100 percent, compared to neat plasma or serum proteomics without depletion or enrichment. The novel ENRICH-iST workflow enables fast and robust processing of up to 96 plasma samples in parallel for reproducible large-cohort translational and clinical research and validation. The affordable ENRICH-iST routine enrichment workflow nicely complements existing ultra-deep plasma proteomics discovery research workflows that either use automated bead depletion or nanoparticle array enrichment.

The ENRICH-iST technology offers a simple, fast (<5h total) and cost-effective solution for proteomic sample processing of plasma or serum from various species, including humans, rodents, NHPs, dogs, and pigs. Requiring only 20 microL of starting material, it consistently delivers reproducible, enhanced results for LC-MS analysis, e.g., by DIA-PASEF timsTOF 4D Proteomics mass spectrometry. Utilizing non-functionalized paramagnetic microbeads, the technology enables biologically unbiased enrichment of lower abundant proteins and supports liquid handling automation platforms, such as Tecan FLUENT systems.

Dr. Andreas-David Brunner, Drug Discovery Sciences, Boehringer Ingelheim Pharma GmbH & Co. KG, states: “We routinely work with biofluid cohorts collected from preclinical species. The novel ENRICH technology from PreOmics allowed us to achieve reliable and in-depth proteomic data without laborious method development. The new kit worked out of the box; the workflow is intuitive and can be completely automated. This solution allows us to standardize our processes completely, and significantly improves our throughput.”

In a benchmark study, plasma and serum samples from human, mouse, and rat specimens (from suppliers Sigma Aldrich, BioIVT, and Diaserve Laboratories GmbH) were processed using the novel ENRICH-iST workflow in comparison to antibody-based depletion, as well as to unprocessed neat samples. Peptides were analyzed on a timsTOF HT mass spectrometer (Bruker Daltonics) in DIA-PASEF mode using a 30-minute nanoLC gradient. The ENRICH-iST workflow was shown to increase the proteomic depth of peptides and proteins 1.5x – 2.2x -fold in comparison to neat samples. Typical Coefficients of Variation (CV) within replicates were below 12 percent, emphasizing high repeatability of the ENRICH-iST workflow. Matching the identified proteins with the plasma proteome database revealed a superior coverage for the ENRICH-iST workflow over neat and depleted samples with identified proteins covering a larger dynamic range. ENRICH-iST showed higher precision over the measured abundance range in comparison to the depletion workflow.

Dr. Catherine Nury, Manager Proteomics, Philip Morris International, Switzerland, comments: “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 are always there to support us. We would strongly recommend the kit to other plasma and serum proteomics users.”

Automation is key for high-throughput plasma proteomics and Luca Valeggia, Senior Vice President—Lab Automation, at Tecan, adds: “We are delighted to continue our collaboration with PreOmics. By automating their new ENRICH-iST kit on our Fluent workstation, we’re enabling robust and reproducible high-throughput sample preparation for large cohort studies, helping to further accelerate progress in this rapidly advancing field.”