In this work, a highly efficient profiling of N-linked glycans was achieved by a facile and eco-friendly synthesized highly porous metal-free carbon material. The metal-free carbon was derived from a well-defined nanorod zinc metal-organic framework via the metal removal under a high-temperature carbonization, which exhibited a highly specific surface area of 1700 m/g. After further oxidation, the oxidized metal-free carbon was applied to the selective isolation of N-linked glycans from complex biological samples due to the strong interaction between carbon and glycan as well as the size-exclusion mechanism. Twenty six N-linked glycans could be identified from the digest of a standard glycoprotein ovalbumin at a concentration of 0.01 μg/μL, and the detection limit of glycans could be down to 1 ng/μL with 21 N-linked glycans identified. When the mass ratio of the interfering protein bovine serum albumin vs a standard ovalbumin digest is up to 500:1, there were 24 N-glycans confidentially identified. From a real complex sample of a healthy human serum, there were 43 N-linked glycans identified after the enrichment of oxidized metal-free carbon. In a word, the metal-free carbon is opening up new prospect for the high-throughput identification of glycan.
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http://dx.doi.org/10.1021/acsami.8b02423 | DOI Listing |
Talanta
December 2024
Institute of Analytical Chemistry of the Czech Academy of Sciences, Veveri 97, 602 00, Brno, Czech Republic. Electronic address:
In this work, we present the synthesis and application of fluorescent rhodamine B hydrazide for the derivatization of simple oligosaccharides and complex glycans using a hydrazone formation chemistry approach. The labeling conditions and the experimental setup of CE/LIF were optimized by analyzing oligosaccharide standards. The CE/LIF separations were performed in polybrene-coated capillaries eliminating the need for the purification step after derivatization.
View Article and Find Full Text PDFBiotechnol Bioeng
December 2024
Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, Delaware, USA.
The CHO VRC01 cell line produces an anti-HIV IgG1 monoclonal antibody containing N-linked glycans on both the Fab (variable) and Fc (constant) regions. Site-specific glycan analysis was used to measure the complex effects of cell culture process conditions on Fab and Fc glycosylation. Experimental data revealed major differences in glycan fractions across the two sites.
View Article and Find Full Text PDFSci Rep
December 2024
Department of Respiratory Medicine, Hunan Provincial People's Hospital (The First-Affiliated Hospital of Hunan Normal University), No. 61 Jiefang Xi Road, Changsha, Hunan, 410219, China.
Pulmonary arterial hypertension (PAH) is a serious medical condition that causes a failure in the right heart. Two-pore channel 2 (TPC2) is upregulated in PAH, but its roles in PAH remain largely unknown. Our investigation aims at the mechanisms by which TPC2 regulates PAH development.
View Article and Find Full Text PDFMethods Mol Biol
December 2024
Department of Biochemistry, Center for Biomedical Mass Spectrometry, Boston University School of Medicine, Boston, MA, USA.
Mass spectrometry-based investigation of the heterogeneous glycoproteome from complex biological specimens is a robust approach to mapping the structure, function, and dynamics of the glycome and proteome. Sampling whole wet tissues often provides a large amount of starting material; however, there is a reasonable variability in tissue handling prior to downstream processing steps, and it is difficult to capture all the different biomolecules from a specific region. The on-slide tissue digestion approach, outlined in this protocol chapter, is a simple and cost-effective method that allows comprehensive mapping of the glycoproteome from a single spot of tissue of 1 mm or greater diameter.
View Article and Find Full Text PDFbioRxiv
December 2024
Department of Biochemistry, University of Washington, Seattle, WA.
A critical step in infections is the attachment of many microorganisms to host cells using lectins that bind surface glycans, making lectins promising antimicrobial targets. Upon binding mannosylated glycans, FimH, the most studied lectin adhesin of type 1 fimbriae in , undergoes an allosteric transition from an inactive to an active conformation that can act as a catch-bond. Monoclonal antibodies that alter FimH glycan binding in various ways are available, but the mechanisms of these antibodies remain unclear.
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