The purpose of this pilot study was to investigate the effects of the transfusion of one erythrocyte concentrate on the number of circulating red blood cell extracellular vesicles (RBC‐EVs) and their clearance time. Six, healthy volunteers donated their blood and were transfused with their RBC concentrate after 35–36 days of storage. One K2EDTA and one serum sample were collected before donation, at four timepoints after donation and at another six timepoints after transfusion. RBC‐EVs were analyzed on a Cytek Aurora flow cytometer. A highly significant increase (p < 0.001) of RBC‐EVs from an average of 60.1 ± 19.8 (103/μL) at baseline to 179.3 ± 84.7 (103/μL) in the first 1–3 h after transfusion could be observed. Individual differences in the response to transfusion became apparent with one volunteer showing no increase and another an increased concentration at one timepoint after donation due to an influenza infection. We concluded that in an individualized passport approach, increased RBC‐EVs might be considered as additional evidence when interpreting suspicious Athletes Biological Passport (ABPs) but for this additional research related to sample collection and transport processes as well as method development and harmonization would be necessary.
The current study examined the stability of several antidoping prohibited substances analytes in urine after 15‐min exposure to UV‐C light in a Biosafety Level 2 cabinet. The urine matrices were exposed within the original antidoping bottles with the aim to destroy DNA/RNA and possible SARS CoV‐2. The analytes small molecules Phase I and Phase II metabolites and peptides, in total 444, endogenous, internal standards, and prohibited substances, pH, and specific gravity in urine were studied. The accredited analytical methods were used by Anti‐Doping Laboratory Qatar for the comparison of data of the same urine samples analyzed with and without UV‐C exposure. In the study conditions, no problems of stability were detected in the substances spiked in the urine samples exposed in the UV‐C irradiation.
Antidoping testing for recombinant human erythropoietin (EPO) is routinely performed by gel electrophoresis followed by western blot analysis with primary and secondary antibodies. The two antibody steps add more than 24 h to the testing time of a purified sample. The aim of this study was to test the concept of using directly horseradish‐peroxidase (HRP)‐conjugated anti‐EPO primary antibody, without the need for a secondary antibody, to reduce the analysis time and eliminate non‐specific cross‐reactivity with secondary antibodies. An in‐house, periodate coupling (R&D systems, clone AE7A5) and three commercially available anti‐human EPO‐HRP conjugates from Genetex, Novus Biologicals and Santa Cruz were evaluated for specificity and sensitivity, using recombinant human EPO standards, negative human urine samples and urine samples from an EPO excretion study. The in‐house anti‐EPO‐HRP conjugate was performed as well as the current two‐step application of unconjugated primary and secondary antibodies used in routine analysis, with comparable specificity and sensitivity. The analysis time was markedly reduced for purified samples from 25 h with the routine method down to 7 h with the in‐house HRP conjugate. Of the three commercially available conjugates tested, only the Santa Cruz anti‐EPO‐HRP conjugate showed comparable specificity but had lower sensitivity to both the in‐house and the antibody combination currently applied routinely. The other two commercially available conjugates (Genetex and Novus Biologicals) did not show any visible bands with the EPO standards. The results clearly demonstrate the potential utility of a directly HRP‐conjugated anti‐EPO antibody to reduce analysis time for EPO in doping control.
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