Abstract
Changes in the abundance of antennary fucosylated glycans in human total plasma N-glycome (TPNG) have been associated with several diseases ranging from diabetes to various forms of cancer. However, it is challenging to address this important part of the human glycome. Most commonly, time-consuming chromatographic separations are performed to differentially quantify core and antenna fucosylation. Obtaining sufficient resolution for larger, more complex glycans can be challenging. We introduce a matrix-assisted laser desorption/ionization—mass spectrometry (MALDI-MS) assay for the relative quantitation of antennary fucosylation in TPNG. N-linked glycans are released from plasma by PNGase F and further treated with a core fucosidase before performing a linkage-informative sialic acid derivatization. The core fucosylated glycans are thus depleted while the remaining antennary fucosylated glycans are quantitated. Simultaneous quantitation of α2,3-linked sialic acids and antennary fucosylation allows an estimation of the sialyl-Lewis x motif. The approach is feasible using either ultrahigh-resolution Fourier-transform ion cyclotron resonance mass spectrometry or time-of-flight mass spectrometry. The assay was used to investigate changes of antennary fucosylation as clinically relevant marker in 14 colorectal cancer patients. In accordance with a previous report, we found elevated levels of antennary fucosylation pre-surgery which decreased after tumor resection. The assay has the potential for revealing antennary fucosylation signatures in various conditions including diabetes and different types of cancer.
Highlights
Changes in the relative abundance of either core or antennary fucosylation have been associated with certain diseases or disease states (Blomme et al, 2009; Testa et al, 2015)
We developed an assay for the relative quantitation of antennary fucosylation in human total plasma N-glycome (TPNG) by combining MALDIMS and exoglycosidase approaches
Completeness of core defucosylation was judged by the inability to quantify by MALDI-MS (signal to noise ratio (S/N) < 9) some of the most abundant core fucosylated glycans namely [H3N4F1 + Na]+ of m/z 1,485.53 and [H4N4F1 + Na]+ of m/z 1,647.59 (Supplementary Figure S4)
Summary
Changes in the relative abundance of either core or antennary fucosylation have been associated with certain diseases or disease states (Blomme et al, 2009; Testa et al, 2015). The abundance of antennary fucosylation in TPNG has been correlated with (1) certain cancers such as hepatocellular carcinoma (Benicky et al, 2014; Zhu et al, 2014), ovarian cancer (Saldova et al, 2007) and colorectal cancer (Holst et al, 2016; de Vroome et al, 2018; Doherty et al, 2018); (2) hepatocyte nuclear factor 1 homeobox A—maturity-onset diabetes of the young (HNF1A—MODY) (Thanabalasingham et al, 2013; Juszczak et al, 2019); (3) inflammatory conditions (Brinkman-van der Linden et al, 1998; Higai et al, 2005) and (4) with attentiondeficit hyperactivity disorder in children (Pivac et al, 2011). Quantitation of antennary fucosylation in TPNG has been found to be an HNF1A-MODY disease biomarker with potential for complementing genetic tests in disease diagnosis (Lauc et al, 2010; Thanabalasingham et al, 2013) For colorectal cancer, both better diagnosis and prediction of long-term survival are urgent clinical needs. Antennary fucosylation was shown to be associated with the recovery of colorectal cancer patients after tumor resection (de Vroome et al, 2018), and may have the potential for long-term survival prediction
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
