Olof Beck

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.

Lithium is the first-line treatment for bipolar disorder. However, the narrow therapeutic window of serum (s-)lithium is near its toxicity range, necessitating continuous monitoring of patients, a process involving regular hospital visits. On-demand home sampling could allow for more frequent testing, possibly resulting in safer patient outcomes, further dosage optimization, and increased compliance. This article presents a device that measures the s-lithium concentration from whole blood. The device consists of a single-use cartridge able to conduct on-chip serum filtration, volume-metering and an on-chip colorimetric assay. Spiked whole blood shows good linearity (Pearson’s r = 0.96, R² = 0.92), a limit-of-detection of 0.3 mmol/L, and an average deviation of 0.05 mmol/L (± 6 %) compared to atomic absorption spectroscopy. The on-chip colorimetric assay has shown to be a promising technique for measuring s-lithium concentration from whole blood and could allow patients to assess lithium levels at home and make the treatment available for new patient groups.

The recent COVID-19 pandemic has posed major health challenges. To expand our molecular understanding beyond those seeking medical care, we conducted a cross-sectional survey targeting random residents from Sweden’s two largest cities. Two thousand participants were invited during 2021 to self-sample dried blood spots (DBS) and provide health information. DBS samples were analysed for multiple anti-SARS-CoV-2 antibodies (Abs), auto-reactive antibodies (AAbs) against 23 interferons (IFNs), and for 502 circulating immune-related proteins. Data-driven methodologies characterised the DBS phenotypes in response to infections and vaccination. We received DBS samples from 437 (22%) volunteers aged 18-69, 60% females, and 50% per city. Multi-analyte serology distinguished self-reported infections (26%) and vaccination (40%), revealing a time-dependent discrepancy between reported and measured immunity. Anti-IFN Abs were detected in 21% of the donors and more frequent for type 1 IFNs alongside a natural infection. Integrating proteomics with antibody data provided additional insights into processes of cell-mediated immunity. Proteomics-centric analysis identified 24% of the participants to deviate in their phenotypes due to infection, immunity, respiratory distress, and age-related traits. Multi-molecular analysis of layperson samples uncovered heterogeneity and diversity of immunity and health phenotypes, complementing clinical investigations.

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 (LHB), follicle-stimulating hormone (FSHB), thyroid-stimulating hormone (TSHB), prolactin (PRL), and growth hormone (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.

Illicit drug use is common among attendees of electronic dance music (EDM) festivals, but is often significantly underreported by participants. The current study aimed to compare the prevalence and over- and under-reporting of illicit drug use among attendees at EDM festivals in two European countries with distinct drug laws and cultures. Self-reported data regarding recent drug use were collected through interviews. Participants’ blood alcohol concentrations were measured using a breathalyzer. Recent illicit drug use was assessed through sampling microparticles in the breath and consequent off-site analysis through liquid chromatography and tandem mass spectroscopy. Illicit drug use was higher in Belgium than in Sweden as indicated by self-reports (56.8 vs. 4.3%) and drug testing (37.2 vs. 12.5%). Underreporting was higher in Sweden than in Belgium; in Sweden, only 2.6% reported taking an illicit drug other than cannabis, whereas 11.6% tested positive, while the corresponding figures in Belgium were 36.5% and 36.9%. In both countries, results from self-reporting and drug testing for specific drugs matched poorly at the individual level, indicating unwitting consumption of substances. This study indicates that the drug use prevalence and the likelihood of disclosure may differ between countries or cultures, which should be considered when choosing methods to investigate drug use prevalence.

Phosphatidylethanol (PEth) measurement in whole blood samples is established as a specific alcohol biomarker with clinical and forensic applications. Establishment of dried blood spots (DBSs) as a specimen for PEth determination offers several advantages and was the focus of this work. A liquid chromatography tandem mass spectrometry method using a 96‐well format for sample preparation was developed and validated. PEth was extracted from DBSs by using isopropanol containing PEth‐d5 as internal standard. The blood sampling used a commercial volumetric DBS device having a phosholipase D inhibitor incorporated to stop continuous PEth formation. The method quantified PEth in the range of 0.05–10 μmol/L, with a bias and imprecision of less than 15%. In a clinical study (n = 25) using fingerprick blood, the volumetric device offered more precise quantifications (CV 4.6%) compared with the Whatman 903 Protein Saver card device (CV 16.6%). In another clinical study (n = 48), the use of dried venous and capillary blood, and liquid venous blood was compared under real‐life conditions with samples sent by postal service. The capillary and venous DBS samples gave identical results while the liquid blood gave slightly higher values. Calculation of elimination half‐life (PEth 16:0/18:1) in 31 cases based on two consecutive samples with 2–9 days in between gave results (mean 6.2 days) that agree with literature but several cases with values over 10 days. In conclusion, this study demonstrates that volumetric DBS is a valid specimen for determination of PEth blood concentrations, offering several advantages.

This phase II clinical trial investigates a one-time oromucosal dose of tetrahydrocannabinol/cannabidiol (THC/CBD) in 23 patients with indolent leukemic B cell lymphomas. Primary endpoint was a significant reduction in leukemic B cells. Grade 1 − 2 adverse events were seen in 91% of the patients; most common were dry mouth (78%), vertigo (70%), and somnolence (43%). After THC/CBD a significant reduction in leukemic B cells (median, 11%) occurred within two hours (p = .014), and remained for 6 h without induction of apoptosis or proliferation. Normal B cells and T cells were also reduced. CXCR4 expression increased on leukemic cells and T cells. All effects were gone by 24 h. Our results show that a single dose of THC/CBD affects a wide variety of leukocytes and only transiently reduce malignant cells in blood. Based on this study, THC/CBD shows no therapeutic potential for indolent B cell lymphomas (EudraCT trial no. 2014-005553-39).