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Journal Article Describes Optimized On-Bead Digestion Method


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Biotech Support Group reports on a recently published journal article describing an improved method for on-bead digestion of serum proteins first enriched with AlbuVoid™. The citation is:

Zheng H, Zhao C, Qian M, Roy S, Arpa A, et al. (2015) AlbuVoid™ Coupled to On-Bead Digestion - Tackling the Challenges of Serum Proteomics. J Proteomics Bioinform 8: 225-230. doi:10.4172/jpb.1000373

In brief, the article describes new methods using a surface/bead based product, AlbuVoid™, which depletes Albumin through a negative selection or voidance strategy, retaining the vast amount of the remaining serum proteome on the bead.

Then the enrichment is combined with a direct and seamless integration with trypsin digestion, a method conventionally referred to as on-bead digestion. The digest time was evaluated as a parameter to identify whether different sub-populations of peptides and proteins can be observed by LC-MS analyses.

Using 2 different allotted digestion times - 4 hours, and overnight, each with a singular 3 hour gradient LC-MS run, between 400-500 total proteins were observed for both human and rat sera, with overlapping and distinct sub-populations observable at each digest time. These results support that the described methods gain efficiencies over other high abundance depletion and in-solution digestion workflows.

“We are pleased with these developments as serum can be a particularly difficult sample to work with in proteomics. With these improved and efficient on-bead methods, depletion and digestion of serum and plasma samples will become more robust and reproducible. Furthermore, the method can be generally applied for discovery proteomics, with either label or label-free quantitation.” states Swapan Roy, Ph.D., President and Founder of Biotech Support Group.

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