Research Article Cites NuGel™ PBA for Proteome Analysis of Saliva
The authors describe the influence of two different sample pre-treatments on the detection of stress biomarkers in saliva. Both the enrichment of glycoproteins by immobilized boronic acid (PBA) and sub-proteome enrichment by dynamic range compression were considered in this study. For this purpose, saliva samples collected before and after the application of an acute stress model in pigs were evaluated. Two-dimensional gel electrophoresis (2-DE) followed by MALDI-TOF/TOF mass spectrometry (MS) was used as the proteomic reporting technique. The article states “1mg of total protein from each salivary pool was lyophilized and subjected to PBA treatment. A commercial glycoprotein enrichment kit based on PBA matrix was used (NuGel™ Glycoprotein enrichment PBA kit, Biotech Support Group) according to the manufacturer’s instructions.” In brief, “the freeze-dried PBA saliva was dissolved in 250μL of binding buffer, added to 50mg of PBA matrix and incubated for 10min. The unbound fraction was discarded…the matrix was washed three times with 350μL of wash buffer, and the glycoprotein-enriched fraction was obtained by incubation of the matrix with 300μL of sorbitol elution buffer…”. The authors concluded that the use of different procedures allowed the detection of different potential stress biomarkers. From a practical point of view, the use of saliva without treatment as well as the enrichment of glycoproteins with NuGel™ PBA were preferred. Read More
Discovery Of Glycoprotein Biomarkers Using NuGel™ Glycoprotein Enrichment Phenyl Boronic Acid (PBA) Kit.
Providing unrivalled specificity, NuGel™ Glycoprotein Enrichment PBA Kit contains phenyl boronic acid which covalently reacts with 1,2-cis-diol group–containing molecules. This unique chemistry eliminates the requirement for forming complex saccharide recognition motifs for interaction. The PBA ligand binds to the 1,2-cis-diol groups of biomolecules and enriches for heterogeneous sets of glycoproteins containing both N-linked and O-linked oligosaccharides. The specialized heterogenous nature of NuGel™ Glycoprotein Enrichment PBA Kit’s organo-boronic acid groups makes it applicable to all kinds of glycopeptides regardless of the sizes, structures, and hydrophilicities. The boronic acids facilitate the capturing of heterogeneous groups of O-linked oligosaccharides.
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