E-Lynx Newsletter

The National Center for Functional Glycomics (NCFG), with the support of the NIH Common Fund for Glycosciences, will be screening a selection of Vector Labs' lectins.  Using the Consortium for Functional Glycomics (CFG) glycan arrays, the NCFG analyzes the glycan specificity of each of our lectin lots, and provides these detailed lectin-glycan binding data on the NCFG website. The goal of this NCFG project is to support rigorous and reproducible glycobiology experiments.

These data provide uniquely valuable insights into glycan structures on the glycoproteins you are studying:

• Binding specificity of each lectin, for hundreds of glycan structures in the array
• Data tabulated and conveniently summarized as a binding array, with peak height indicating degree of binding for each glycan
• Specific and detailed data on carbohydrate structure
• Lot-specific data—for confidence in the reproducibility of your binding data

There are times when chemical transfection may not be optimal for certain applications. Electroporation is a great solution for hard-to-transfect cell types like immune or stem cells or to deliver diverse molecules such as DNA, RNA or protein.

Click here to watch. (Link to YouTube)

Detection of endotoxin that may be present in biopharmaceutical products is critical to patient safety.  Although the endotoxin molecule itself is highly stable, various factors such as matrix composition, storage temperature and container makeup can affect its stability in manufacturing samples that are collected and subsequently assayed for endotoxin content. Pfizer in Andover, MA developed a procedure for creating an in-house naturally occurring endotoxin (NoE) preparation to assess the stability of endotoxins across various matrices, temperatures and containers.  The use of NoE provides benefits over using commercially available Control Standard Endotoxin (CSE), such as increased laboratory flexibility and control, and has been used by Pfizer to assess endotoxin stability over time in various matrices and temperatures.

Allegro™ Targeted Genotyping allows interrogation of DNA heterogeneity at an industrial scale.  By using NuGEN’s patented Single Primer Enrichment Technology (SPET) we provide strand-specific readouts of genome sequence, enabling sensitive detection of allele frequency for as many as hundreds of thousands of sites.  Our custom panel design process is a complimentary concierge service that will provide designs that meet the specific business or scientific needs of your study.  Targeted sequence data are amenable to industry-standard genotyping analysis pipelines.  Additionally, we provide the ability to perform genotyping through a cloud analysis platform built for Allegro™ customers.  Here, we present a validation study using an oligo panel targeting human SNPs.  Reference gDNA with a known allele mixture is used as input into Allegro™; we observe high concordance with known minor-allele frequencies ranging from 5% to 35% (pearson R > 0.98).  Our process grants significant cost-reductions compared to whole-genome assays at a price-point comparable to or better than microarray approaches with the fidelity and scalability benefits of Next Generation Sequencing.

Nucleosome substrates are emerging tools for the in vitro study of chromatin epigenetic regulation, with applications from basic chromatin biology research to high-throughput drug screening. The advent of recombinant designer nucleosomes (dNucs) carrying fully defined covalent modifications of the histone proteins (e.g. lysine acylation) or wrapping DNA (e.g. 5’ methylcytosine) enables novel or dramatically improved approaches (e.g. reader binding, enzymatic assays, antibody profiling) for next generation chromatin research.

In this post we are going to talk about the minimal thresholds for the assembly and quality validation of these reagents.

Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease worldwide.  Even though underlying mechanisms are not fully understood, pathways responsible for the activation of inflammatory cytokines like TNF and members of the IL-1 cytokine family are key in the development of NAFLD. Recent studies reported that not only the inflammasome-caspase-1 pathway is responsible for the activation of proinflammatory cytokines: neutrophil serine proteases (NSPs) are as well.  This opinion article describes new insights about the NSPs as potent cytokine activators that contribute to liver disease progression.

Why Alpha-1-antitrypsin?
Alpha-1-antitrypsin (AAT) is a natural occurring serine peptidase inhibitor produced by the liver of both inflammasome-dependent and independent cytokine activation pathways.  It is the most abundant circulating protease inhibitor known.  It mainly targets enzymes released by neutrophils, especially neutrophil elastase (NE) but also proteinase 3 (PR3) and Cathepsin G (CG).  These NSPs are stored in the azurophilic granules of neutrophils.  When neutrophils are activated, NSPs will be released and can activate for example proinflammatory cytokines.  In the extracellular matrix AAT can inhibit NSPs and this leads to the inhibition of cytokine activation.  A deficit of AAT is responsible for insufficient inhibition of NSPs and thus for inflammation-induced liver diseases.  In patients, AAT has already been shown to be a successful therapy in COPD and cystic fibrosis: Administration of AAT in patients with COPD or cystic fibrosis has proven efficient in inhibiting neutrophil elastase activity in the lung function.  Since AAT is a potent inducer of IL-1Ra it is a reason to consider that this protein is a good candidate for treating IL-1-mediated diseases.

Summary of Mirea, A et al; IL-1 Family Cytokine Pathways Underlying NAFLD: Towards New Treatment Strategies. Trends in Molecular Medicine 2018; Volume 24, Issue 5, Pages 458-471 . 

Click here for the full article. (Link to NCBI website)

Ion exchange chromatography separates molecules based on the overall charge of the protein.  The proteins of interest have a charge opposite to that of the resin. In the case of cation exchange chromatography, proteins have an overall positive charge while binding to a cation exchanger.  The initial binding takes place under low ionic strength conditions. Elution is achieved by a salt gradient.  By increasing the salt concentration, proteins with a weak positive charge elute first, while at higher salt concentrations, proteins with a strong positive charge elute later.  Ion exchange chromatography is frequently used for protein purification.  Precise gradient formation is an important parameter for the successful separation of  proteins.  AZURA Bio Lab System supports all gradient methods including ion exchange chromatography.

Taking a walk may be a good opportunity to mentally review your to-do list, but using the time to instead be more mindful of your breathing and surroundings may help boost your wellbeing, according to researchers who found that while students reported being less stressed while they were on their feet and moving, they received an even greater benefit when they reported also being more mindful.

Click here to read more.  (Link to ScienceDaily website)

How Meditation Saved the Thai Soccer Team Trapped in a Cave

"Moments of crisis can trigger fight-or-flight responses for even the most mindful and centered folks. That's why meditation was key for the Wild Boars, the Thai soccer team that was rescued from a cave this week after catastrophic flooding."

Click here to read more.  (Link to MindBodyGreen website)

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