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Multiplex quantification of endocrine proteins in volumetric dried blood spots
KTH, Centres, Science for Life Laboratory, SciLifeLab. KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Protein Engineering.
Department of Laboratory Medicine, Clinical Chemistry, Östersund Hospital, Östersund, Sweden; Department of Medical Sciences, Clinical Chemistry, Uppsala University, Uppsala, Sweden; Capitainer AB, Solna Torg 19, Solna, Sweden, Solna Torg 19.
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Affinity Proteomics. KTH, Centres, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0001-9329-2353
Department of Clinical Neuroscience, Karolinska Institutet, Stockholm, Sweden.
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2025 (English)In: Clinical Proteomics, ISSN 1542-6416, E-ISSN 1559-0275, Vol. 22, no 1, article id 18Article in journal (Refereed) Published
Abstract [en]

Background: Circulating proteins are routinely quantified from liquid biopsies to deduce health and disease. Among these are endocrine protein hormones, which regulate human growth, development, metabolism, and reproduction. Most commonly, these proteins are analyzed in plasma or serum prepared from venous blood draws. Recently, devices for quantitative capillary sampling from a finger prick have emerged, but their utility for clinical testing remains to be explored. Methods: To study the analytical capabilities of quantitative dried blood spots (qDBS), we quantified the luteinizing hormone subunit beta (LHB), follicle-stimulating hormone subunit beta (FSHB), thyroid-stimulating hormone subunit beta (TSHB), prolactin (PRL), and growth hormone 1 (GH1) by multiplexed immunoassays. We determined the performance of the endocrine hormone assays in paired qDBS and EDTA plasma samples from 100 donors (90% females) aged 4 to 78. Lastly, we compared the protein levels with those from an accredited clinical chemistry laboratory. Results: The multiplexed analysis showed precise protein quantifications in qDBS (mean CV = 8.3%), high concordance with plasma levels (r = 0.88 to 0.99), and accuracy being matrix- and protein-dependent (recovery: 80–225%). Using the current protocol and sample dilutions, reported protein concentrations were 1.2 to 7.5 times higher in plasma than in qDBS eluates. Concentrations from multiplexed plasma assays agreed with the clinical data (r = 0.87 to 0.99) and decreased slightly when comparing clinical plasma data with multiplexed qDBS assays (r = 0.76 to 0.98). Significant increases in age-related FSHB and LHB levels were observed in females in all specimens and assays (p < 0.01). Conclusions: This study shows the suitability of modern qDBS devices for quantifying clinically informative proteins in multiplexed assays and highlights the need for future work on specimen-specific optimization and standards. Volumetric DBS sampling offers new routines for accurate protein quantification for precision medicine.

Place, publisher, year, edition, pages
Springer Nature , 2025. Vol. 22, no 1, article id 18
Keywords [en]
Dried blood spots, Endocrine hormones, Multiplexed immunoassays, Quantification, Women’s health
National Category
Endocrinology and Diabetes
Identifiers
URN: urn:nbn:se:kth:diva-363796DOI: 10.1186/s12014-025-09539-3ISI: 001485492300001PubMedID: 40346485Scopus ID: 2-s2.0-105004676561OAI: oai:DiVA.org:kth-363796DiVA, id: diva2:1959892
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QC 20250526

Available from: 2025-05-21 Created: 2025-05-21 Last updated: 2025-06-02Bibliographically approved

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Bendes, AnnikaSchwenk, Jochen M.

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