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NRicher™ Serum/Plasma Knowledgebase
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Introduction. Quantitative serum/plasma proteomics can help unravel regulatory elements contributing to disease. Yet, extracting and characterizing functional changes and adaptations to disease for many of even the highest abundance proteins in circulation remains limited. Some reasons are an over-emphasis on proteome coverage demanding Venn Diagram comparisons of analytical platforms and methods, along with a lack of cost-effective and scalable targeted proteomic workflows. The latter is in part due to the changing landscape of proteins/peptides not associated with the selected targets, oftentimes called analytical matrix effects, or background noise. The NRicher™ Knowledgebase & Supporting Products/Methods provides a solution. Researchers can find their protein(s) of interest, and determine the NRicher™ bead/methods to best enrich for those targets. Its free to review, and accessible on a non-confidential basis. Over 2000 proteins, from a pooled normal/healthy population are annotated with corresponding relative signal intensities for each bead/method. Additional annotations include curated prospective biomarkers, soluble membrane proteins, and the interconnected sub-proteomes of innate immunity. NRicher™ target(s) enrichment can minimize acquisition time, collectively improving overall throughput, with outstanding gains in productivity. NRicher™ methods offer scalability, automation compatibility, and cost-effectiveness, without any upfront instrument requirements
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Purpose: Researchers can determine whether pre-identified protein targets of interest can be enriched by one or more NRicher™ beads; user defined targets may come from sources such as:
a. Discovery proteomics
b. Gene Expression
c. Curated from public domain databases and publications

Function of the Knowledgebase
Over 2000 proteins observed with corresponding relative signal intensities
Find your protein(s) of interest, and corresponding bead/methods to best enrich for protein(s) of interest, free to review, accessible on a non-confidential basis
Annotation of 200 soluble membrane proteins, derived from ectodomain shedding, a common dysregulated disease mechanism, and important source of precision biomarkers
Annotation of 100+ prospective biofluid biomarkers associated with disease conditions, noting the optimal products for enrichment
New strategies to compartmentalize and profile systemic chronic inflammation, and associated diseases

Why Choose NRicher™
For over 10 years BSG has been at the forefront of developing synthetic beads (i.e., ionic, hydrophobic, hydrogen bonding, aromatic, polymeric) with differential proteome binding properties.
The NRicher™ Advantage
Not derived from immuno-affinity: NRicher™ beads, are not species-specific. This allows a wider applicability across various sample types.
Cost-efficient:
There's no need for an investment in high-end specialized equipment; a standard laboratory microfuge will suffice.
Streamlined Analysis:
Through the use of bead cocktails, NRicher™ products serve virtually all applications, starting with sample volumes as low as 25 µl.
On-bead digestion:
BSG pioneered Bead-Assisted Sample Prep (BASP™), offering workflow efficiencies (i.e., no added denaturants) for LC-MS proteomics of bead enriched sub-proteomes.
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Coming Soon Use the Knowledgebase for New Functional Proteomic Strategies To Profile Innate Immunity, Systemic Chronic Inflammation and Associated Diseases
Chronic inflammation, and its association to disease, is strongly linked to the triangle of interconnected pathways of coagulation, complement and neutrophil recruitment. Indeed, the amplitude, and temporal dynamics of proteases and their inhibitors are critical factors influencing disease.
Yet, conventional proteomic investigations on innate immunity proteins are often misleading, as common methods only produce static measurements, assuming a direct correlation of abundance to function. However, characterizing the innate immunity proteome is not so direct, as the innate response is activated through proteolytic mechanisms. As as result, strict static measurements can be egregiously unreliable. The Knowledgebase offers new categorical strategies to help characterize these critically important functionalities.
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Proteomic biomarker profiles can characterize immune dysregulation with applications in early disease detection, risk evaluation, prognosis, and patient stratification. Many of the biomarker proteins listed mirror responses associated with acute phase inflammation, innate immunity (i.e., Complement), coagulation and degradation of platelets, tissue remodeling (soluble membrane proteins), as well as neutrophil & macrophage heterogeneity/polarization and related granulocytic cargo release.
In addition to the “Biomarker” and “Membrane” Protein Tabs, in future updates to this knowledgebase, we will assign Tabs to categorical proteins according to:
• Complement Activation and Regulation
• Coagulation Proteins and Platelet proteins, categorized by dense and alpha granules; granule secretion being pivotal to establishing and controlling the microenvironment at the local inflammatory site
• Neutrophil cargo release from 5 different granule types in coordinated response to inflammatory insults; sub-proteomes are categorized
• Nine major inhibitory Serpins (i.e., Alpha-1-Antitrypsin, Antithrombin III) account for 5-10% of the protein mass in serum, affirming their importance to maintain normal homeostasis, and coordinating central control for innate immunity.
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The NRicher™, product line continues to advance sub-proteome enrichment, with products/methods that can be tailored to specific applications. Choose Best NRicher™ Enrichment Products for Your Biomarkers of Interest. Here’s How.
Tabs are for different Batches of LC-MS proteome data, each Batch reveals different columns, each column has its own unique bead/method combination. The numbers reflect signal intensities so relative quantities can be evaluated for each protein by row. Contact sales@biotechsupportgroup.com for decoding the column(s) and selection of the best bead/methods for the biomarker(s) of interest. Relative signal levels can be evaluated by ratios of target signal to either total signal or high abundance (i.e., Albumin) signal, or co-eluting peptide signal(s) associated with spectral interferences.
Other Tabs provide additional context to the Knowledgebase, including: > 200 Soluble Membrane Protein derived from ectodomain shedding, a rich family for biomarkers >Disease Associated Biomarkers - 100+ with annotated references for many diseases including cardiovascular, oncology & neurodegenerative, about half are soluble membrane proteins
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A selection of one or more NRicher™ beads can be customized to meet the application requirements. Beyond the selection of bead chemistries, further optimization can be achieved through process protocols, such as load and bind/wash buffer adjustments.

After NRicher™, target peptides have enriched spectral signal, even as gradient times are reduced
NRicher™ sub-proteome enrichment can minimize acquisition time, collectively improving overall throughput, cost, and outstanding gains in productivity
Specific target peptides that report functional and gene variant regions promise actionable insights and potential multiplex biomarkers for disease.
NRicher™ methods offer scalability, automation compatibility, and cost-effectiveness. 		Serpins are a very unique protein family of protease inhibitors, sometimes called suicidal inhibitors, display a decoy trapping mechanism. Upon cleavage at the Reactive Center Loop (RCL), the SERPIN spontaneously refolds into a hyperstable conformation, where the N-terminal portion of cleaved RCL is inserted between central β-sheet A, “trapping” the covalently linked protease into an inactive form. However, SERPINs can also act as substrates for the protease. This results in an active protease that disassociates from the SERPIN, which leaves the SERPIN as a cleaved inactive sub-form. As proteolysis is irreversible, its regulation by Serpins is essential to coordinate and ultimately resolve the innate immune response. Ask about new strategies to functionally monitor this vitally important regulating protein family by LC-MS.
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All NRicher products have the same price structure and protocols based on 25-100 µl serum/plasma per prep: 10 preps, $625 : 50 preps, $2,430. For all inquiries about decoding the products and methods, and for knowledgebase updates, please contact sales@biotechsupportgroup.com
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