---
title: "N-glycome Profiling of Dried Blood Spots for Pre-diabetes and Diabetes Distinction"
id: "biorxiv-0-n-glycome-analysis-of-dried-blood-spots-from-different-blood-preparations-and"
canonical_url: "https://medichelpline.com/clinical-feed/biorxiv-0-n-glycome-analysis-of-dried-blood-spots-from-different-blood-preparations-and"
content_type: "clinical_feed_article"
specialty: "Endocrinology"
source_name: "bioRxiv (Biomedical Preprints)"
source_url: "https://www.biorxiv.org/content/10.64898/2026.08.24.746065v1?rss=1"
published_at: "2026-08-25T09:19:42.000Z"
evidence_level: "Verified Feed"
license: "CC-BY-NC-4.0 / Informational Use"
---
# N-glycome Profiling of Dried Blood Spots for Pre-diabetes and Diabetes Distinction
## Provenance & Clinical Metadata
- **Canonical URL:** https://medichelpline.com/clinical-feed/biorxiv-0-n-glycome-analysis-of-dried-blood-spots-from-different-blood-preparations-and
- **Specialty:** [Endocrinology](https://medichelpline.com/clinical-feed/endocrinology.md)
- **Primary Source:** bioRxiv (Biomedical Preprints)
- **Source URL:** [Original Journal Publication](https://www.biorxiv.org/content/10.64898/2026.08.24.746065v1?rss=1)
- **Published At:** 2026-08-25T09:19:42.000Z
- **Evidence Rating:** Verified Feed
## Executive GIST (TL;DR)
- The study evaluates N-glycosylation profiling from **dried blood spots (DBS)** using an established HILIC-UHPLC-FLR workflow adapted for DBS samples. - The protocol includes DBS cutting, protein extraction and enzymatic digestion, labeling with 2-aminobenzamide, cleanup, and measurement by **HILIC-UHPLC-FLR**. - DBS prepared from fresh blood, frozen whole blood, or separated frozen blood cells with corresponding frozen plasma produced a stable N-glycosylation profile, indicating sample flexibility and stability. - Comparison between DBS and plasma was performed; the method enables N-glycan analysis from nonstandard or repurposed samples such as frozen whole blood. - In comparisons of DBS N-glycans from pre-diabetic and diabetic subjects, trends were observed but did not reach statistical significance: increases in **fucosylation**, **bisection**, and **galactosylation**, and a decrease in **sialylation** in diabetes. - The authors highlight DBS N-glycan profiling as easier, cheaper, and less invasive than conventional plasma sampling for diagnostics and monitoring in pre-diabetes and diabetes. - The approach has potential to repurpose samples not originally intended for N-glycan biomarker analysis, expanding usability for retrospective and resource-limited settings. - Competing interests: G.L. is founder/owner of Genos Ltd; I.T-A. is an employee; E.M. was employed by Genos Ltd during the work. Funding: E.M. supported by an EU Horizon 2020 Marie Skłodowska-Curie grant; funder was not involved in study conduct or publication decisions. - This work is a preprint and has not undergone peer review; specific quantitative details, cohort sizes, statistical values and full methodological parameters were reported in the source but are not expanded in this summary.
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Elham Memarian 1 Genos Glycoscience Research Laboratory, Zagreb, Croatia, Center for Proteomics and Metabolomics, Leiden University Medical Center, the Netherlands; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Elham%2BMemarian%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Memarian%20E&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3AElham%2BMemarian%2B) * [ORCID record for Elham Memarian](http://orcid.org/0000-0001-5862-4375 "Open in new tab") * For correspondence: el1.work.memarian@gmail.com Irena Trbojevic Akmacic 2 Genos Glycoscience Research Laboratory, Zagreb, Croatia; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Irena%2BTrbojevic%2BAkmacic%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Trbojevic%20Akmacic%20I&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3AIrena%2BTrbojevic%2BAkmacic%2B) * [ORCID record for Irena Trbojevic Akmacic](http://orcid.org/0000-0003-0106-0155 "Open in new tab") Ozren Polasek 3 Department of Public Health, University of Split School of Medicine, Soltanska 2, 21000 Split, Croatia 4 Algebra University, Gradiscanska 24, 10000 Zagreb, Croatia; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Ozren%2BPolasek%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Polasek%20O&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3AOzren%2BPolasek%2B) Gordan Lauc 4 Genos Glycoscience Research Laboratory, Zagreb, Croatia, Faculty of Pharmacy and Biochemistry, University of Zagreb, Zagreb, Croatia * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Gordan%2BLauc%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Lauc%20G&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3AGordan%2BLauc%2B) * [ORCID record for Gordan Lauc](http://orcid.org/0000-0003-1840-9560 "Open in new tab") * [Abstract](https://www.biorxiv.org/content/10.64898/2026.08.24.746065v1)[](https://www.biorxiv.org/panels_ajax_tab/biorxiv_tab_art/node:5726968/1) * [Info/History](https://www.biorxiv.org/content/10.64898/2026.08.24.746065v1.article-info)[](https://www.biorxiv.org/panels_ajax_tab/biorxiv_tab_info/node:5726968/1) * [Metrics](https://www.biorxiv.org/content/10.64898/2026.08.24.746065v1.article-metrics)[](https://www.biorxiv.org/panels_ajax_tab/article_tab_metrics/node:5726968/1) * [ Preview PDF](https://www.biorxiv.org/content/10.64898/2026.08.24.746065v1.full.pdf+html)[](https://www.biorxiv.org/panels_ajax_tab/biorxiv_tab_pdf/node:5726968/1) ![Loading](https://www.biorxiv.org/sites/all/modules/contrib/panels_ajax_tab/images/loading.gif) ## Abstract Dried blood spot (DBS) sampling is becoming a popular alternative to traditional blood sampling approaches, offering advantages such as convenience of collection, transportation, and storage, as well as lower biohazard risk. N-glycosylation, a major post-translational modification of proteins associated with numerous biological and pathological functions, is one area of interest for DBS analysis. In this study, we utilize a protocol for N-glycosylation profiling of DBS by ultra-high-performance liquid chromatography based on hydrophilic interactions and fluorescence detection (HILIC-UHPLC-FLR). The protocol includes DBS cutting, protein extraction and enzymatic digestion, labeling with 2-aminobenzamide, followed by cleanup and HILIC-UHPLC-FLR measurement. We compare DBS with plasma and demonstrate the stability of DBS N-glycosylation profile when DBS are prepared from fresh blood, frozen whole blood, or a combination of separated frozen blood cells and corresponding frozen plasma. Additionally, we compared DBS N-glycans from pre- and diabetic subjects. Fucosylation, bisection, and galactosylation showed a statistically non-significant increasing trend in diabetes, whereas sialylation showed a statistically non-significant decreasing trend in diabetes. The main advantage of this method is the ability to repurpose samples, which were initially not intended for biomarker N-glycan analysis, such as frozen whole blood. Additionally, DBS N-glycan profiling is the easier, cheapest and the least invasive approach to conventional plasma in pre-diabetes and diabetes patients' diagnostics and monitoring. ### Competing Interest Statement . G.L. is the founder and owner of Genos Ltd, a private research organization that specializes in high-throughput glycomic analysis and has several patents in this field. I.T-A. is an employee of Genos Ltd. E.M. was employed by Genos Ltd at the time of this work; No other potential conflicts of interest relevant to this article were reported. ## Funder Information Declared E.M. was supported by funding from the European Union’s Horizon 2020 research and innovation program under the Marie Sklodowska–Curie grant for the project GlySign (contract no. 722095). The study funder was not involved in the design of the study; the collection, analysis, and interpretation of data; writing the report; and did not impose any restrictions regarding the publication of the report. Copyright The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission. bioRxiv and medRxiv thank the following for their generous financial support: > The Chan Zuckerberg Initiative, Cold Spring Harbor Laboratory, the Sergey Brin Family Foundation, California Institute of Technology, Centre National de la Recherche Scientifique, Fred Hutchinson Cancer Center, Imperial College London, Massachusetts Institute of Technology, Stanford University, The University of Edinburgh, University of Washington, and Vrije Universiteit Amsterdam. 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