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Glycosphingolipids (GSLs) are glycolipids with ceramide and carbohydrate head groups that play an important role in numerous biological processes. Previously, we performed GSL-glycan analysis of various cell lines and virus-infected cells using a glycoblotting approach. Recently, we developed several methods for sialic acid linkage-specific chemical modification to distinguish sialylated glycan isomers by mass spectrometry. In this chapter, we describe a method for analyzing GSL-glycans in human serum/plasma using glycoblotting combined with aminolysis-SALSA (sialic acid linkage-specific alkylamidation) and lactone-driven ester-to-amide derivatization (LEAD)-SALSA for comprehensive and detailed structural glycomics.
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http://dx.doi.org/10.1007/978-1-0716-2910-9_21 | DOI Listing |
Glycobiology
September 2025
Department of Chemistry, Maynooth University, Maynooth, Co. Kildare W23 F2H6, Ireland.
Changes in glycosylation can serve as markers for rare genetic disorders, including lysosomal storage diseases (LSDs). Nephropathic Cystinosis (NC), caused by mutations in the CTNS gene, is characterised by cystine accumulation in lysosomes due to dysfunctional cystinosin, a heavily N-glycosylated lysosomal transporter. We analysed total serum and IgG N-glycosylation using hydrophilic interaction ultra performance liquid chromatography (HILIC-UPLC) to explore the diagnostic biomarker capabilities and their pathophysiological relevance in NC.
View Article and Find Full Text PDFNeurol Res Pract
August 2025
Department of Neurology, Rechts Der Isar Hospital of the Technical University Munich, Munich, Germany.
Introduction: The median time to diagnosis of amyotrophic lateral sclerosis (ALS) is approximately 12 months after the onset of first symptoms. This diagnostic delay is primarily due to the nonspecific nature of early symptoms and the clinical challenges in differentiating ALS from its mimics. Therefore, the discovery of reliable biomarkers for the early and accurate diagnosis of ALS represents a critical medical need.
View Article and Find Full Text PDFJ Pharm Biomed Anal
December 2025
Bioanalytical Sciences, Genentech Inc, 1 DNA Way, South San Francisco, CA 94080, USA. Electronic address:
Immuno-polymerase chain reaction (iPCR) is an analytical technology that combines the specificity of antibody reagents with the sensitivity of polymerase chain reaction (PCR) signal amplification. Here we describe the optimization and qualification of an ultra-sensitive iPCR method designed to detect and quantify active (reduced form) interleukin-33 (IL-33) in human samples. The modified assay incorporates several improvements over a previous version utilized in research studies.
View Article and Find Full Text PDFOrphanet J Rare Dis
August 2025
Board of Directors & Staff, Association for Creatine Deficiencies, Carlsbad, CA, USA.
Background: Creatine transporter (CTD) and guanidinoacetate methyltransferase (GAMT) deficiencies are rare inborn errors of creatine metabolism, resulting in cerebral creatine deficiency. Patients with either condition commonly exhibit intellectual and developmental disabilities, often accompanied by behavior problems, delayed speech, seizures, and motor impairments. There is currently no efficacious treatment for CTD, while current management for GAMT requires lifelong treatment with a protein restricted diet and intake of high amounts of oral supplements.
View Article and Find Full Text PDFJ Chromatogr A
September 2025
Institute of Environmental Assessment and Water Research (IDAEA-CSIC). Barcelona, Catalonia, Spain. Electronic address:
Human biomonitoring of persistent organic pollutants (POPs) remains essential for tracking long-term exposure, evaluating health risks, and assessing the effectiveness of regulatory bans. For this purpose, an experimental and analytical methodology has been optimized allowing the determination of 47 POPs, encompassing chlorinated cyclodienes, chlorobenzenes, cyclohexanes, polychlorodiphenyl derivatives, several congeners of polychlorobiphenyls (PCBs) and polybromodiphenyl ethers (PBDEs, including the deca-BDE), as well as three non-persistent chemicals (namely quintozene, tecnazene and vinclozolin). The method uses a single liquid-liquid extraction procedure with 500 µL of serum/plasma.
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