Functional & Top-down Proteomics

Building sequence/function relationships

The continuum of protein conformations attributable to post-translational modification and non-covalent interactions produces characteristic functions. Thus, functional annotation complements sequence annotation, but relies in part, on the functional or structural features of intact, non-denatured proteins. BSG has developed a new method for proteome separations based on chemically derived 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

The related NRicher™ (6 & Mx) products are adaptable to any measurable protein function from any sample, to aid in the discovery of new biomarkers. They support all functional, activity-probe, chemical and top-down proteomic applications. So functional protein attributes, as when the same or similar underlying sequence can have multiple conformations and functions, or when different sequences cross-over in function, are now open to investigation.


NuGel™ NRicher™ 6

Functional proteomics and enrichment kit

  • 12 differentiated subproteomes, 6 flow-through fractions, and 6 elution fractions
  • Uncompromised functional and structural integrity
  • Enrich low abundance functional biomarkers for sequence and structural annotation
  • In a rigorous examination of protein complexes, about twice the number of observations were made possible through sub-proteome bias characteristics of NRicher™ 6 { Nature doi:10.1038/nature14877 }
  • Kit includes 6 mixed mode bead chemistries per prep
  • All Top-down proteomic applications
  • Optimize biomarker enrichment after initial enrichment using NRicher™ Mx
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NuGel™ NRicher™ 6

PEP Functional Activity Analysis, courtesy of ArrayBridge (St. Louis, MO). Each well was monitored for Hexokinase activity using beef extract for cascade enzymes; NADP reduction being the final reporting measurement. The circled regions are activities up-regulated after subtraction of background. The observed activities show a different pattern for each of the 6 NRicher™ beads – designated A,B,C,L,N,R, and generally a feature pattern distinguishing the normal (upper panel) from the colon cancer (lower panel) sera.

NuGel™ NRicher™ Mx

Low Abundance Proteome Enrichment

  • Enriches & normalizes sub-proteomes
  • Compress proteome concentrations
  • Species and tissue agnostic
  • Composite of the NRicher™ 6 mixed mode beads
  • Suitable for small volume mg scale preps
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NRicher™ Mx Eluates

Non-reduced SDS-PAGE profiles show that major high abundance bands are greatly reduced. The low abundance bands are enriched relative to their high abundance bands, visually estimated to be 5-10X for most bands. Because of its heterogeneity, the IgG variants co-localize to the same region.


Our BSG Advantage


Consumable

Cost-effective, not derived from biologicals

Cost-effective, not derived from biologicals

  • No specialized instruments or HPLC
  • Economical surface chemistries, not derived from biologicals
  • No regeneration, so no prep to prep variability
  • Simple, fast microfuge bind/wash/elute protocols

On-Bead Digestion

Efficient workflows, quality LC-MS/MS data

Efficient workflows, quality LC-MS/MS data

  • Simple, reproducible workflows
  • Equivalent or better than in-solution digestion
  • Seamless to LC-MS, no desalting or C18 separations
  • Unique proteolytic efficiencies

Enrichment/Depletion

Diverse strategies, species agnostic

Diverse strategies, species agnostic

  • Products support strategies for both enrichment of low abundance proteomes, or depletion of high abundance proteins
  • Species agnostic not derived from biologicals

Functional Integrity

Maintained throughout all separations

Maintained throughout all separation

  • Mild buffer conditions maintains native structure with retained enzymatic, functional & bio-activities
  • Supports enzyme biomarker assays
  • Functional & Chemical Proteomics
  • Structural & activity-probe Proteomics
  • Top-down & ArrayBridge PEP Proteomics