Glycans in Therapy and Diagnostics

A special issue of Bioengineering (ISSN 2306-5354). This special issue belongs to the section "Biochemical Engineering".

Deadline for manuscript submissions: closed (15 December 2022) | Viewed by 1818

Special Issue Editors


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Guest Editor
ProBioGen AG, Berlin, Germany
Interests: glycosylation; vectored vaccines

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Guest Editor
Department of Engineering, HTW-Berlin University of Applied Sciences, 12459 Berlin, Germany
Interests: bioprocess engineering; carbohydrate interactions; cell line engineering; glycan engineering; glycobiology; intracellular N-glycan processing; sugar nucleotide metabolism; therapeutic glycoproteins
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Guest Editor
Department of Life Sciences and Technology, Beuth University of Applied Sciences, 13353 Berlin, Germany
Interests: biochemistry of glycans; chemical biology; glycan engineering; glycan analysis; glycobiotechnology; sialic acids; therapeutic glycoproteins

Special Issue Information

Dear Colleagues,

Over the last two decades, glycans have evolved as relevant diagnostic biomarkers and also as a quality attribute for manufactured therapeutic glycoproteins. Glycans are a major constituent of glycolipids as well as secreted and cell surface glycoproteins and account for the overall structural microheterogeneity of these molecules. Human-type glycan structures are characterized by a set of individual glycan moieties composed of varying numbers of glycosidically linked hexose and desoxyhexose units and neuraminic acid. Aberrant patterns of glycosylation have been observed in the context of several diseases and are therefore being developed into valuable biomarkers that help to distinguish a healthy from the diseased state and lift the power of diagnostic assays to yet another level. The heterogenic profile of N- and O-glycan compositions within a manufactured batch of bulk drug substance affects the overall efficacy of the biologic drug and is therefore considered to be a critical quality attribute that needs to be monitored throughout production and in the context of product release. The industry has started to address this need to match specific desirable glycan compositions by glycan engineering of host cells as well as by linking process parameters to drug substance glycan quality attributes via QbD type process design and development accompanied by concomitant monitoring of the drug substance glycan structure. Glycan discovery, glycan engineering and glycan analytics are among the most important fields of current biopharma research. There is an urgent need for advancement in all of these sectors.  This Special Issue of Bioengineering on “Glycans in therapy and disease” comprises expert contributions from industry and academia on glycans in the context of diagnostic biomarker discovery and development, glycan structure as a target for molecular engineering of production hosts for therapeutic glycoproteins and current advances in glycan analytical technology. 

Dr. Volker Sandig
Prof. Dr. Hans Henning Von Horsten
Prof. Dr. Stephan Hinderlich
Guest Editors

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Published Papers (1 paper)

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Research

13 pages, 3074 KiB  
Article
Seamless Coupling of Chemical Glycan Release and Labeling for an Accelerated Protein N-Glycan Sample Preparation Workflow
by Mumtaz Kasim, Anja Griebel, Grit Sandig, Robert Höltzel, Akshay Malhotra, Stephan Hinderlich, Volker Sandig, Barbara Müller and Hans Henning von Horsten
Bioengineering 2023, 10(6), 651; https://0-doi-org.brum.beds.ac.uk/10.3390/bioengineering10060651 - 26 May 2023
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Abstract
Analytical methods fr direct quantitative N-glycan analysis require a sequence of sample preparation and clean-up steps that result in reduced glycan recovery. Therefore, we aimed to combine glycan release and labeling steps. Based on the hypothesis that the reaction mechanism for oxidative [...] Read more.
Analytical methods fr direct quantitative N-glycan analysis require a sequence of sample preparation and clean-up steps that result in reduced glycan recovery. Therefore, we aimed to combine glycan release and labeling steps. Based on the hypothesis that the reaction mechanism for oxidative chemical glycan release comprises a stable glycan isocyanate intermediate, we investigated whether this could be exploited for the in-situ preparation of fluorescent glycan conjugates. ANTS-labeled N-glycans were derived from chicken ovalbumin via an in-situ chemical release/coupling approach and by standard Peptide-N-Glycosidase F (PNGase F) digestion/reductive amination. Synoptic fluorescence-assisted carbohydrate electrophoresis with UV detection (FACE-UV) analysis yielded matching patterns of fluorescent N-glycan bands in the expected electrophoretic mobility range between hexose units GU-5 and GU-11 of the standard. Anthranilamide (2-AB)-glycan conjugates prepared from a test glycoprotein carrying a predominant Core-F glycan gave single predominant peaks in hydrophilic interaction chromatography with fluorescence detection (HILIC-FLD) and electrospray ionization mass spectrometry (ESI-MS) spectra in agreement with sodiated triply charged Core-F-AB conjugates for both the standard and the in-situ coupling methods. The Core-F-AB conjugate prepared by the in-situ coupling approach had a slightly elevated retention time on HILIC-FLD and an ESI-MS m/z peak in line with a urea-bonded glycan-AB conjugate, with closed pyran ring structures on the glycan moiety. Glycan isocyanates intermittently formed during chemical glycan release, which could be utilized to prepare labeled glycan samples directly from glycoproteins and fluorescent dyes bearing a primary amine functional group. Full article
(This article belongs to the Special Issue Glycans in Therapy and Diagnostics)
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