Sample Prep for Mass Spectrometry

  • Proteome Enrichment
  • Not based on immuno-affinity
  • Bead Assisted Sample Prep (BASP™)
  • LC-MS, MALDI Compatible Protocols

High abundance proteins and lipids are well known interferences in mass spectrometry based proteomic analysis. For years the protein depletion toolkit was limited primarily to immuno-affinity chromatography and other biologically-derived tools.

BSG offers two complementary technologies for proteomic researchers to selectively bind or enrich, in order to achieve the best results. By using consumable non-biologically derived beads, researchers will be able to improve efficiency and quantitation essential for expanding mass spectrometry into routine healthcare.


AlbuVoid™ LC-MS On-Bead

Albumin depletion plus low abundance protein enrichment, coupled with optimized on-bead digestion (BASP™) protocols for LC-MS serum and plasma proteomics

  • Seamless workflows
  • Unique proteolytic efficiencies
  • Label, label free compatible
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Total LC-MS/MS Proteins identified at 2 digestion times


Total LC-MS/MS Proteins identified at 2 digestion times: 4 hours (4h) & overnight (O/N), single 3 hour LC gradient.

AlbuVoid™ PLUS

Albumin and IgG Depletion From Serum/Plasma for Proteomics

  • IgG removal >90% (70-80% of total Immunoglobulins removed)
  • Albumin removal >95%
  • Seamless and simple < 1 hour protocol
  • Low abundance enrichment equivalent to immuno-affinity
  • Disposable, cost-effective, no column regeneration or cross-contamination
  • Works for most species tested including human, sheep, rat, mouse, bovine
  • On-bead protocols improve workflow and efficiency, especially suited to targeted proteomics
  • Suitable for LC-MS, 1 and 2D Gels, ELISAs, Enzyme and other Functional Assays.
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SDS-PAGE: Comparison of three methods using Human Serum Sample


20µg of protein loaded on to 10% tris bis gel

A - Serum alone – shows IgG and Albumin regions
B - NuGel™ ProteinA treated serum – shows IgG depletion
C - AlbuVoid™ treated serum – shows Albumin depletion
D - AlbuVoid™ PLUS treated serum – shows IgG and Albumin depletion

HemoVoid™ LC-MS On-Bead

Hemoglobin depletion plus low abundance protein enrichment, coupled with optimized on-bead digestion protocols (BASP™) for LC-MS erythrocyte & blood proteomics

  • Seamless workflows
  • Unique proteolytic efficiencies
  • Label, label free compatible
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Total Proteins Identified From Erythrocytes

AlbuSorb™

Selectively Binds Albumin

  • Removes serum albumin >90%
  • Economical small ligand surface architecture (not dye-based), bio-affinity performance
  • Species agnostic
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AlbuSorb™ PLUS

Selectively Binds Albumin & Immunoglobulin

  • > 85% Albumin, 85% IgG depleted from 25 μl serum
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Quantitative Bias Comparison between AlbuSorb™ & AlbuSorb™ PLUS

AlbuSorb™ AlbuSorb™ PLUS
Total IG IDs 2+ Spectral Cts 80 58
Total IG % of Spectral Count 24% 9%
Total Albumin % of Spectral Count 12% 15%
Total Proteins IDs

HemogloBind™
and
NuGel™ HemogloBind™

Removes Hemoglobin Interference

  • Highly specific for hemoglobin binding
  • depletion from hemolyzed serum, dura, BALF, and whole blood
  • supports biomarker quantitative analysis
View HemogloBind™ View NuGel™ HemogloBind™
HemogloBind™

Cleanascite™

Lipid Adsorption & Clarification

  • Effectively replaces chlorinated/fluorinated hydrocarbons (eg. freon)
  • Workflows for antibodies, proteins, nucleic acids, proteoglycans, and most serum analytes
  • A high binding capacity for lipids with minimal cross-reactivity with proteins and nucleic acids
  • Ideal for clarifying ascites, serum, cell & tissue culture, bile and organ homogenates
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Cleanascite™ is supplied as a suspension reagent.

Cleanascite™ is supplied as a suspension reagent. Simply add, mix and centrifuge in a 10 minute protocol.


References

Plasma/Serum Protein Biomarkers

For diabetes biomarkers, Cleanascite™ is shown both to improve LC-MS measurements, and validated in accordance with CLIA ’88 guidelines. { doi: 10.1016/j.cca.2016.01.019 }


Bile Proteomics

Cleanascite™ treatment was indispensable for in-solution digestion methods. { https://pubag.nal.usda.gov/catalog/5599484 }


For MALDI-TOF Analysis

Farina A, Dumonceau JM, Frossard JL. Proteomic Analysis of Human Bile from Malignant Biliary Stenosis Induced by Pancreatic Cancer Journal of Proteome Research.2009; 8(1):159-69

NuGel™ NRicher™ 6

Top-down proteomics enrichment kit

  • 12 differentiated subproteomes, 6 flow-through fractions, and 6 elution fractions
  • 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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LC-MS Protein Identifications
NRicher™ 6 Improved dynamic range and ID coverage by sub-proteome capture Nature doi:10.1038/nature14877

With NRicher™ 6 Beads and Without

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
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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