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Using AlbuVoid™ as a Serum Protein Enrichment Kit in Functional Proteomics


Validation of an albumin depletion and low abundance protein enrichment kit, AlbuVoid™, for negative selection or voidance of albumin with low abundance enrichment of remainder sub-proteome from serum, plasma, tissues and culture media.

Background

For years the protein depletion toolkit was limited primarily to immuno-affinity chromatography and other biologically-derived tools. While effective for many applications, such tools are not efficient for “omics” sample preparation, as throughput, economy and simplicity are especially required. Furthermore, these same separation tools often denatured proteins which limited there use in applications which demanded the measurement of function, structure or bio-activity. For these reasons, BSG is dedicated to create new methods and applications to drive efficient workflows and better data quality for all proteomic and biomarker analyses.

BSG offers a unique variety of general non-specific beads, or stated another way - beads with weak affinity or imperfect fit interactions. Without the use of antibodies, progressive displacement allows the beads to bias for or against certain proteins, without compromising protein integrity. Consequently, our products find many diverse uses to profile non-invasive body fluids and liquid biopsy sample types such as blood, saliva, synovial fluid and urine. Because of our unique advantages, seamless integration with analytical platforms such as LC-MS, ELISA/Western Blot, and 2DE are in common use.

Furthermore, with the many advances in functional proteomics, our products are now being cited in journal articles such as “ Identification of potential serum biomarkers for breast cancer using a functional proteomics technology”. The full citation is:

David L. Wang, Chuanguang Xiao, Guofeng Fu, Xing Wang and Liang Li. Identification of potential serum biomarkers for breast cancer using a functional proteomics technology. Biomarker Research (2017) 5:11. DOI 10.1186/s40364-017-0092-9

The Challenge

Over the past decade, proteomic comparisons have been applied for discovery of potential biomarkers from human body fluids, including blood and serum/plasma. For cancer, these biomarkers are expected to have great utility for screening and early detection, treatment response evaluation, and recurrence monitoring.

With new technologies now available to identify and validate functional proteins as biomarkers, it’s essential to do so without compromising the quality or quantity of the biomarkers present. To achieve a high level of sensitivity and specificity in detection of functional biomarkers, low abundance protein enrichment during the discovery process can be a key consideration.

The Solution

In this particular study cited above, the authors’ used both enriched serum samples and compared those to neat serum pools (without enrichment). In this study, a functional proteomics technology was used to systematically monitor metabolic enzyme and protease activities from resolved serum proteins produced by a modified 2-D gel separation and subsequent Protein Elution Plate, a method collectively called PEP (Array Bridge, St. Louis, MO). Sera from breast cancer patients and normal/healthy people were compared.

As a pre-treatement enrichment, AlbuVoid™ was shown to enrich enzyme activity for subsequent analysis. As a result, additional candidate fractions become available to choose from for further biological validation as potential biomarkers.

The article states “The most dramatic difference for enzyme activity detection in using the AlbuVoid for serum protein enrichment was demonstrated. … Compared with the direct serum proteinase measurement, both the levels and species of proteases were increased significantly in the enriched serum sample. …protease activity in the direct serum analysis suggested that the protease levels in the serum were below the detection threshold of protease activity…,and it is necessary to use AlbuVoid to enrich these low level proteases to bring them to a high enough level to be detected.”

The Outcome

A key feature in all of our products is to not only separate proteomes but to retain the functional features of proteins after separations. Without a reliable serum protein enrichment kit like AlbuVoid™, the study could have been compromised because the low level of protease activities would not have been brought to a high enough level sufficient for detection.

This article, along with the citations below, illustrate the importance of simple and efficient proteomic sample preparation technology to retain the functional features of proteins after separations. AlbuVoid™ and our full product line is designed to help profile proteome conformations that are highly variable, each conformation contributing to its own unique functional activity.

Key Advantages of AlbuVoid

  • Albumin voids in flow-through >95%, with <30 minute bind/wash/elute protocol
  • Disposable, cost-effective, no column regeneration or cross-contamination
  • Species agnostic

View Albunim & IgG Removal Kits

Notable References that feature AlbuVoid™

View all of our references

BSG Featured Advantage: Functional Integrity

Functional integrity maintains post-separations and supports functional, chemical and top-down proteomics. The mild buffer conditions maintain native structure with retained enzymatic, functional & bio-activities.

Notable References that feature the Functional Integrity Advantage

  • Cortese-Krott, Miriam M., et al. "Identification of a soluble guanylate cyclase in RBCs: preserved activity in patients with coronary artery disease." Redox Biology (2017).

http://www.sciencedirect.com/science/article/pii/S2213231717306535