If you’re involved in plasma sample preparation, you’re probably already aware of the difficulties that come along with it. Blood plasma is frequently used in biomarker discovery due to its accessibility and richness of information about a person’s health status. However, the high-dynamic range of proteins, combined with the sample complexity, makes accessing low-abundance proteins very challenging. In addition, outcomes are less robust and reproducible due to complex and time-consuming workflows. Furthermore, efficient automation to screen in a manageable time frame larger cohorts, for example in the context of drug discovery approaches, poses quite a challenge to current workflows.
In this post, we outline the limitations of traditional plasma proteomic techniques, and share a new approach that finally reduces drastically the high dynamic range in plasma and serum samples, as well as streamlines your workflow significantly.
To solve the dynamic range challenge for efficient high-throughput plasma proteomic analyses, various techniques have been developed. Methods such as fractionating plasma samples or enriching specific groups of proteins can enhance our understanding of the plasma proteome beyond what unprocessed plasma reveals. However, they require extensive hands-on and measurement time.
Conventional depletion of high-abundance proteins, such as albumin and immunoglobulins can also reveal previously masked low-abundance proteins. Yet, using immunodepletion kits or columns is often labor-intensive and unsuited for high-throughput techniques. Immunoaffinity-based depletion is specific to each species, limiting the potential sample types. This method also risks potential cross-reactivity of antibodies or co-depletion of proteins alongside carrier proteins like albumin, which affects the accuracy of identifying target proteins.
The key to the dynamic range challenge of plasma samples is PreOmic’s complete ENRICH-iST workflow. This user-friendly yet robust solution utilizes the biologically unbiased enrichment of low-abundance proteins onto paramagnetic particles (EN-BEADS), reducing the dynamic range while conserving analytical depth and proteome coverage. Another core component of the ENRICH-iST’s efficacy comes from PreOmic’s iST-BCT approach, which is an automatable, three-step protocol optimized for biological fluids in terms of alkylation rate and minimal artificial modifications. The protocol also allows for high-throughput processing of up to 96 samples in parallel and is compatible with human, mouse, and rat samples. With exceptional reproducibility and low CVs, the process takes only 5h from sample to pure peptides.
To learn more about the materials and methods behind ENRICH-iST, please view our Application Note.
A clear, efficient, and automatable workflow for preparing plasma and serum samples is crucial for conducting reliable research, such as in cohort studies. In particular, improved analysis of liquid biopsy samples is vital for gaining deeper insight into pathogenic disease mechanisms and is crucial for biomarker discovery.
Enter PreOmics, an industry leader in providing innovative tools for mass spectrometry-based proteomics. Our innovative ENRICH-iST kit provides an easy-to-use, yet robust solution to the dynamic range challenge of plasma and serum samples.
If you’re interested in learning how to increase your sample throughput, improve quality, and produce reliable data in the time that it takes to get from Boston to Chicago, let’s start a conversation today.
“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.” Catherine Nury, Manager Proteomics, Philip Morris International, Switzerland
“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.” Hyonson Hwang, Ph.D., Research Scientist, Caris Life Sciences
“It 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.” Dr. Roman Fischer, Associate Professor and Head of Discovery Proteomics Facility, University of Oxford
For more testimonials feel free to visit PreOmics’ testimonial page.