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NRicher™ Ig
Enrichment of all isotypes and subclasses of Immunoglobulins
Consumable chemically derived beads, species agnostic as they are not derived from antibodies
Enrich circulating immune complexes from sera or plasma from both animals and humans, >90% Albumin removal
Does not require any specialized instruments, just a standard microfuge
Bead format suitable for automation compatibility, please inquire
On-Bead digestion for LC-MS analysis, or optional elution for any functional, enzymatic, or immunoassay analysis
A comprehensive analysis of the humoral immune response (the immunome) has potential to greatly impact research across numerous fields. For example, serum autoantibodies against tumor-associated antigens have recently emerged as early stage biomarkers for different types of cancers. Most autoantibody profiling work has been based on the reactivity of unbound antibodies towards antigens produced by a variety of strategies (i.e., cDNA libraries, phage display).
An alternative approach is based on the identification of Ig-bound antigens using Liquid Chromatography coupled to Mass Spectrometry (LC-MS). Such determination of antigens complexed with antibodies at a proteome scale is critical to understanding adaptive responses in the context of infection, autoimmunity, and cancer.
Click Here To View NRicher™ Ig Product Sheet
Human serum immunoglobulins comprise several classes IgG, IgA, IgM, IgD & IgE. IgG is the predominant human immunoglobulin class in plasma and comprises four subclasses; ~60% are IgG1, followed by ~30% IgG2, ~7% IgG3 and ~3% IgG4. To date, most of the circulating antibody complex research has been focused on IgG as the efficiency of recovering a representative pool of IgG antibodies is well established. Generally for human serum/plasma, Protein A binds with high affinity to IgG1, IgG2, and IgG4, but poorly to IgG3. Among the four IgG subtypes in mice, Protein A has the weakest affinity for IgG1 while Protein G has affinity for all four IgG subclasses. Neither Protein A or G bind particularly well towards IgA, IgM, IgD or IgE.
Nevertheless, the ability to enrich circulating immune complexes from sera or plasma from both animals and humans with high yield and without selective loss of isotypes or subclasses can provide more comprehensive profiles. NRicher™ Ig can provide such enrichment for all immunome profiling methods. For antigen reactivity profiling, elution conditions are mild (pH 9-10), and preserve functionality. For antigen identification, bound proteins can be digested on-bead, with seamless integration to LC-MS analysis.
Product
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Size
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Total serum/plasma
samples processed
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Item No.
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NRicher™ Ig
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10 Preps
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10 x (25-50) µl samples
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NIMM-10
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NRicher™ Ig
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50 Preps
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50 x (25-50) µl samples
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NIMM-50
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References
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Swapna LS, Stevens GC, Sardinha-Silva A, Hu LZ, Brand V, Fusca DD, et al. (2024 )
ToxoNet: A high confidence map of protein-protein interactions in Toxoplasma gondii. PLoS Comput Biol 20(6):
e1012208. https://doi.org/10.1371/ journal.pcbi.1012208
The article states we used affinity beads (NuGel PROspector) to pre-enrich Toxoplasma gondii lysate to capture five distinct
subproteomes. [Note: NuGel PROspector beads are now part of the NRicher™ platform.]. When comparing the 5 different
subproteomes, there is clearly different selection biases amongst the 5 surface chemistries. Also, many of the proteins
observed from the NRicher™ beads, were not observed in the Ion exchange fractions demonstrating the importance of
combining different modes (ionic, hydrophobic, etc.) of separation to alter selection properties, and consequently improving
overall proteome coverage.
Wan, C., Borgeson, B., Phanse, S. et al. Panorama of ancient metazoan macromolecular complexes. Nature 525, 339–344 (2015). hXps://doi.org/10.1038/nature14877
Six different NRicher™ beads (described with an old tradename PROspector) were used as an enrichment step in the overall
workflow; about twice the number of observations and annotations became possible. This further validates that the subproteome bias characteristics of the NRicher™ surface chemistry platform can simplify complex proteomes into enriched subproteomes with efficiencies suitable for deep functional proteome characterization.
Whitepaper - NRicher™: A Low Abundance Proteome Enrichment Platform With Seamless Integration of On-Bead Digestion
The NRicher™ Advantage is described: • Consumable chemically derived NuGel™ beads, species agnostic as they are not
derived from antibodies • Does not require any specialized instruments, just a standard microfuge • Use of bead cocktails
allows for one, rather than multiple LC-MS analyses • Functionally active sub-proteomes after separations, for any
orthogonal functional, enzymatic, or immunoassay analysis
https://www.biotechsupportgroup.com/v/vspfiles/templates/257/pdf/BiotechSupportGroup-NRicher-Whitepaper.pdf
NRicher : Family Specific Enrichment For Targeted Proteomics – Poster US HUPO 2024
The need for new biomarkers to support personalized healthcare, has fostered numerous proteomic innovations. Still, a
number of challenges remain. One is the preponderance of high abundance proteins and, concurrently in targeted proteomic
workflows, efficiency and consistency in quantifying target peptides from different sample cohorts. This is in part due to the
changing landscape of proteins/peptides not associated with the selected targets. A solution for both these challenges is now
available through a suite of products called NRicher .
https://www.biotechsupportgroup.com/v/vspfiles/templates/257/pdf/NRicher%20poster%20small.pdf
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