Background
Blood plasma is widely used in proteomics research due to its rich composition of proteins and peptides. It is easily obtainable and can provide valuable insights into various physiological and pathological conditions. As “liquid biopsy,” plasma analysis has become a valuable tool in the context of biomarkers and drug discovery, including cancer and infectious diseases research, as well as drug metabolism.
LC-MS-based proteomics has been extensively used for identifying and quantifying proteins in plasma. However, the high dynamic range (up to 12 orders of magnitude) and the heterogeneity and complexity of plasma samples pose significant challenges for LC-MS analysis, limiting access to the entire proteome.
The Challenge
Accessing low-abundance proteins in plasma and serum samples for LC-MS-based proteomic analysis can be difficult owing to the blood complexity itself and the high-dynamic range of proteins within the biological samples. High-abundance proteins such as albumin and immunoglobulins 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, such as disease diagnosis and drug development.
Researchers at Caris Life Sciences faced the high-dynamic range issue with plasma while
working with human plasma for proteomic mass spectrometry analysis.
Dr. Hyonson Hwang, Research Scientist at Caris Life Sciences, said: “When I first started
working with human plasma for proteomic mass spec analysis, I quickly ran into the well-known dynamic range issue with plasma. I tried several different standard sample preparation protocols but struggled to get adequate results.”
Many labs use standard sample preparation protocols to prepare samples for proteomic
analysis, but scientists often struggle to get fast, reliable, and reproducible results. These
protocols are time-consuming, involving extensive hands-on time, and consequently, more
prone to errors due to variability in sample handling and interference of contaminants, for
example. Likewise, the impracticality of automation, lack of flexibility, and inability to produce
unbiased results impede the application for large sample cohorts.
Expending much time with proteomics sample preparation? Read: Four Ways to Get Better
Sample Preparation Results While Saving Time & Labor.
The Solution
To overcome these challenges, the Caris Life Sciences lab adopted the iST proteomic sample preparation technology. ENRICH-iST delivers a straightforward solution to the dynamic range challenge in blood-derived biofluids by enabling proteins to attach to paramagnetic beads depending on their physical and chemical properties, providing an ideal workflow for the preparation and analysis of small to larger sample cohorts in a wide array of basic and preclinical research.
Dr. Hyonson Hwang added further: “It consistently outperforms any standard methods I used in the market, and I fell in love with it due to the following reasons: overall ease of use, the very helpful color-coordinated reagents and protocol steps, and the consistently reproducible results…Instead of spending time trying to optimize mass spec sample preparation protocols like before, using PreOmics’ products, I have more time for data analysis and interpretation, experimental design, and development of mass spec data collection methods."
Webinar: Discover how to confidently prepare your plasma cohorts with the ENRICH-iST technology in a streamlined workflow for plasma (or serum) profiling and biomarker discovery.
Dr. Roman Fischer, Associate Professor and Head of Discovery Proteomics Facility, University of Oxford, also adopted the PreOmics technology and highlighted its benefits: "[the ENRICH-iST workflow] improved the proteome depth by 4-fold for plasma and 1.6-fold for CSF compared to nondepleted samples. It is surprisingly easy to use, allowing us to prepare 96 samples simultaneously.” His lab combined ENRICH-iST with the Evosep and Tims-TOF Pro platforms to get reliable analysis of large patient cohort sample sets to advance their ongoing clinical and pre-clinical research projects.
Application Note: See how the ENRICH-iST workflow provides an efficient solution for high-throughput proteome profiling of blood-derived biofluidic samples.
Scientists at Boehringer Ingelheim Pharma GmbH & Co. KG routinely work with biofluid cohorts collected from preclinical species and see improvements in their throughput analyses with ENRICH-iST.
Dr. Andreas-David Brunner, R&D Scientist at Boehringer Ingelheim Pharma GmbH & Co. KG, stated: “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 improve our throughput.”
Poster: See how ENRICH-iST can be easily automated.
Meet ENRICH-iST: A streamlined and reliable solution for plasma proteomics
ENRICH-iST is an all-in-one solution involving the enrichment of low-abundance proteins on paramagnetic beads, followed by digestion and purification using the PreOmics iST-BCT protocol. It is compatible with human samples and other mammalian species (e.g., mice, rats, pigs, or dogs). The process from raw samples to pure peptides takes only 5h, and the analysis of large cohorts is possible with established lab automation systems suitable to magnetic bead processing.
If you’re looking for high-performance solutions and workflows that set the standard for protein analysis, let’s start a conversation today.