Case Studies

Proteomic innovationcontinues to drive the need for new biomarkers. For untargeted proteomics, coverage has become an important goal. Nevertheless, for targeted proteomics, efficiency and consistency in quantifying target peptides from different sample cohorts presents signal to noise challenges due to the changing landscape of proteins/peptides not associated with the selected targets.The NRIcher™ bead chemistry platform offers a simple sample prep solution. In this case study, we describe how our Knowledgebase brings advantages to targeted proteomic workflows.

Macrophage polarization refers to how macrophages have been activated as they are sufficiently plastic to integrate multiple signals, such as those from microbe infections, damaged and cancerous tissues, as well as the normal tissue environment. Three BSG products and methods are described in this case study that can help study macrophage polarization.

Through a regulated proteolytic mechanism called ectodomain shedding, the extracellular domains of membrane-anchored proteins are sometimes released from the cell surface as soluble proteins. As a result, any dysregulation within shedding processes can result in diverse pathologies ranging from inflammatory conditions to cancer.

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.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 are annotated with corresponding relative signal intensities for each bead/method. NRicher™ methods offer scalability, automation compatibility, and cost-effectiveness, without any upfront instrument requirements.

Dried blood spot (DBS) analysis offers the advantage of collecting small blood volumes without the need for phlebotomy. The role of DBS analysis by MS now encompasses many other areas of bio-analysis, including and proteomics. We report on simple methods to either deplete Hemoglobin or as in the case of Hemoglobinopathies, isolate Hemoglobin from DBS, for a variety of research purposes.

Red Blood Cells, in addition to oxygen/carbon dioxide exchange, remain at the convergence of all blood-borne processes. While many proteomic efforts have focused on the red cell membrane, less is known about the cytoplasmic fraction, due to the super-abundance of hemoglobin. In this case study, three journal reports describe the utility of HemoVoid™ to deplete Hemoglobin, and help identify potential disease-specific biomarkers from the enriched low-abundance sub-proteome in LC-MS proteomic analysis of red cells and whole blood.

Proteomic studies of the cell secretome have greatly increased in recent years. A major challenge however is the wide dynamic range for the low abundance of genuine secreted proteins, over the highly abundant proteins derived from the serum containing media, the preponderance of which is bovine serum albumin (BSA). In this case study we advance that the secretome can be simply and efficiently depleted of BSA by the product AlbuVoid™.

Lipids are often suspected to induce phenotypic changes in cancer cell lines. However, removing lipids without introducing artifacts that affect cell viability can be problematic. In this case study, we describe the use of Cleanascite™ in published reports to characterize the influence of lipids and other factors bound to lipids, on cell response in cancer.

With the introduction of targeted analysis, sensitivity and technical variance in LC-MS/MS analysis has dramatically improved. Nevertheless, for serum/plasma samples, small biological variances remain hard to measure robustly. In this case study we show how sample prep does matter. By not relying on immuno-affinity for depletion, BSG has strived to address and service the unique demands that come with validating targeted peptide analysis, to accelerate proteomics from discovery to the clinic.

There in increasing evidence associating severe COVID-19 disease with an overly exuberant innate immune response. Our new whitepaper describes how BSG's products and methods can be used to characterize the innate immune response from peripheral blood. Most notably, we present a model of innate immunity which describes a 'protease storm' released by inflammatory cells, which when not sufficiently regulated alters homeostatic balance of proteolysis, which can lead to severe disease.

SARS-CoV-2 (Covid-19) infections utilize membrane bound serine proteases to gain entry into host cells and cause clinical pathology. Endogenous protease inhibitors, especially the Serpin family, also play an important regulating role.

This case study describes our new patent application for the purpose of developing functional Serpin profiles. These will help to monitor the potential for severe disease, as well as the development of new anti-Covid-19 therapeutic strategies.

Red cells are at the convergence of many cellular and extracellular events to which all blood-borne processes come together. Lacking a nucleus, RBCs are unable to synthesize new proteins in response to outside stimuli, making RBCs important reservoirs for biomarker research.

This case study describes many examples of how BSG products efficiently remove hemoglobin from both reticulocyte and mature erythrocyte lysates, enabling such in depth proteomic investigations to flourish.

Unleashing the Power of Proteomics To Better Understand the Innate Immune Response to Infectious and Non-infectious Inflammatory Stimuli

In this case study, we describe how key regulatory elements of the innate immune response can be surveyed by proteomic analysis of serum/plasma, and how proteolysis and its regulation provides the central control function.Thus, the innate immunity proteome can become the source for new biomarkers in the clinical management of COVID-19.

The complex etiology of Parkinson’s disease (PD) is only poorly understood. However, growing evidence now suggests that neurodegeneration in PD is not restricted to dopaminergic neurons in the brain. Rather, PD may be a systemic disease, involving peripheral tissues and may include oxidative, metabolic, or inflammatory processes. So blood based biomarkers may prove useful in early detection of PD as well as to assess the progression of disease in response to medical interventions.

With many products and alternative strategies for depletion and enrichment, Biotech Support Group offers distinct and diverse separations tools that work with virtually all species

Case Study: Methods to monitor the influence and potentially modulate the stromal conditioning of cancer, offers new avenues for diagnosis, personalized medicine, and therapeutic modalities.

Both products deplete Albumin and IgG but in different ways, creating different windows of observation. Evaluating these different windows of observation solves the many challenges of serum proteomics.

Biotech Support Group offers distinct and diverse technologies to help researchers selectively tackle the challenges of serum proteomics, in order to achieve quality results. In this case study, we validate a unique two-step workflow, AlbuVoid™ PLUS, to deplete Albumin and IgG.

In this case study, we highlight two research articles describing Cleanascite™, a solid-phase suspension reagent ideally suited to remove lipids prior to applications involving cell response characterization.

Biotech Support Group offers distinct and diverse technologies to help researchers selectively enrich or bind, in order to achieve quality results. In this case study, we highlight two research articles describing NuGel™ PBA glycoprotein enrichment sample preparation technology for the proteomic analysis of saliva.

Biotech Support Group was recently highlighted in the book, "Functional Proteomics – Methods and Protocols." This authoritative and cutting-edge book, which seeks to fill in the current technology gap, features a collection of technologies developed for the study of protein function at a proteome scale, including much research work from BSG.

By combining two complementary enrichment strategies, Biotech Support Group offers distinctive and diversified technologies to assist researchers achieve quality results by selectively enriching or binding.

Diverse Albumin Depletion and Enrichment Technologies Enhance Efficiency and Simplicity of Achieving Quality Proteomic Information

Validation of a unique Hemoglobin binding reagent, HemogloBind™, for whole blood, dried blood spot and hemolyzed serum.

Two research articles demonstrate useful BSG products for exosome depletion and enrichment needs.

Methods to monitor the influence, potentially modulating the systemic response to cancer, offer new avenues for diagnosis, personalized medicine and therapeutic modalities.

Combining two complementary enrichment strategies, BSG offers diverse technologies for researchers to selectively bind or enrich, in order to achieve the best results.

BSG’s methods development has led to an increased efficiency in protocols used for on-bead digestion of serum proteins, to as fast as two to four hours.

Validation of an albumin depletion and low abundance protein enrichment kit, AlbuVoid™, for negative selection or voidance of albumin with low abundance enrichment of remainder sub-proteome from serum, plasma, tissues and culture media.

Validation of a proteomic sample preparation technology, Cleanascite™, as a simple, yet effective high binding capacity for lipids with minimal cross-reactivity with proteins.

Validation of a commercial resin, HemoVoid, to investigate the efficiency of hemoglobin depletion technology before analysis