According to the World Anti-Doping Agency (WADA) technical document for erythropoiesis stimulating agents (ESA) analysis (TD2014EPO), double-blotting of serum/plasma samples is mandatory for all analysis by isoelectric focusing (IEF) and for the confirmation procedures (CP) performed by SDS-PAGE or SAR-PAGE. The goal is to prevent potential cross-reactions of the secondary antibody with remaining proteins in the purified samples. To this end, we have developed an immunopurification method of ESA in serum/plasma samples using a combination of streptavidin-coated immunomagnetic beads and biotinylated anti-EPO polyclonal antibodies. Here we report that this immunomagnetic bead-based purification allows the analysis of serum/plasma samples by single-blotting. Serum and plasma samples, either intact or spiked with different ESAs, were immunopurified and analyzed by single-blotting, after SAR-PAGE or IEF using a cross-reaction minimized secondary antibody coupled to HRP. The results show that when samples are immunopurified according to this strategy, there is no non-specific binding when single-blotting is performed after SAR-PAGE. With IEF, we observe a faint smearing, however, in the pH gradient outside the ESA detection region. These interferences did not alter ESA profiles of spiked urinary samples or of samples received for routine testing. This approach was compared to the MAIIA monoliths purification or to the isolation of ESAs with other combinations of immunomagnetic reagents (ie, anti-Mouse IgG-coated magnetic beads and anti-EPO mAb). The recovery of ESAs was shown to be significant for serum/plasma samples. Our results suggest that single-blotting could be performed on serum/plasma samples without non-specific interferences. Copyright © 2017 John Wiley & Sons, Ltd.
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http://dx.doi.org/10.1002/dta.2320 | DOI Listing |
Diagnostics (Basel)
December 2024
Department of Drug Chemistry, Pharmaceutical and Biomedical Analysis, Medical University of Warsaw, Banacha 1, 02-097 Warsaw, Poland.
Background: Aldehyde dehydrogenase class 1 (ALDH1) is an enzyme that is ubiquitously distributed in adult tissues and may serve as a prognostic marker in various cancer types. In blood, 99% of ALDH1 is found in erythrocytes; although, it was also demonstrated that leukocytes and platelets exhibit ALDH activity. No ALDH activity was detected in plasma, even when employing the highly sensitive fluorometric method with 7-methoxy-1-naphthaldehyde as a substrate.
View Article and Find Full Text PDFAm J Epidemiol
November 2024
Systems Epidemiology, Faculty of Medicine, University of Oulu, Oulu, Finland.
Mass spectrometry lipidomics is becoming customary to analyse serum/plasma samples in epidemiology. The measurables are molecular constituents of lipoprotein particles, but very little is known on the consequences of adjusting lipidomics data with lipoprotein measures. We studied two population cohorts with 5,657 and 2,036 participants.
View Article and Find Full Text PDFEnviron Int
December 2024
MOE Key Laboratory of Pollution Processes and Environmental Criteria, College of Environmental Science and Engineering, Nankai University, Tianjin 300350, China.
Currently, there is limited knowledge regarding occupational exposure of traditional and emerging organophosphate esters (OPEs) from e-waste and automobile dismantling activities, and their distribution within the human blood. In the present study, we collected dust and urine samples from e-waste (ED) (n = 91 and 130, respectively) and automobile dismantling (AD) plants (n = 93 and 94, respectively), as well as serum-plasma-whole blood samples (sets from 128 participants) within ED areas for analyzing traditional and emerging organophosphate tri-esters (tri-OPEs) and organophosphate di-esters (di-OPEs). Median concentration of ∑tri-OPEs and ∑di-OPEs in dust (37,400 and 9,000 ng/g in ED, and 27,000 and 14,700 ng/g in AD areas, respectively) and urine samples (11.
View Article and Find Full Text PDFInfect Dis Poverty
December 2024
Unidad de Infección Viral e Inmunidad, Centro Nacional de Microbiología - Instituto de Salud Carlos III, Km 2.2, 28220, Majadahonda (Madrid), Spain.
Background: The current diagnostic strategy for hepatitis C virus (HCV) infection involves a two-step approach: antibody HCV screening followed by confirmatory nucleic acid testing. This study aimed to evaluate the diagnostic performance of the Abbott ARCHITECT HCV Ag assay in serum/plasma samples as a potential one-step alternative for diagnosing active HCV infection in people living with hepatitis B virus (PLWHB) through a systematic review and meta-analysis.
Methods: A systematic review and meta-analysis were conducted following PRISMA-DTA guidelines.
Allergy
November 2024
Toulouse Institute for Infectious and Inflammatory Diseases (Infinity), UMR 1291, INSERM, CNRS, University of Toulouse, Toulouse, France.
Background: Diagnosis of allergies is mostly based on the patient's clinical history and allergen provocation tests. Determination of specific IgE (sIgE) profiles can be performed to support allergy diagnosis. This is commonly done in vivo by the skin prick test or in vitro with automated systems.
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