{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,29]],"date-time":"2025-12-29T19:07:00Z","timestamp":1767035220194,"version":"build-2065373602"},"reference-count":36,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2024,9,22]],"date-time":"2024-09-22T00:00:00Z","timestamp":1726963200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Drug Testing and Analysis"],"published-print":{"date-parts":[[2025,7]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>Erythropoiesis\u2010stimulating agents (ESAs) continue to be a significant threat to the integrity of human and equine sports. Besides conventional direct testing, monitoring the biomarkers associated with the effects of ESAs may provide a complementary approach via indirect detection to enhance doping control. In this study, we applied label\u2010free proteomics to discover plasma protein biomarkers in Thoroughbred geldings after administration with a long\u2010acting form of recombinant human erythropoietin (rhEPO), methoxy polyethylene glycol epoetin beta, Mircera. Increased haematocrit, haemoglobin and red blood cell (RBC) levels were evidenced as early as 4\u2009days post\u2010administration in all three horses to varying extents. Tryptic peptides were obtained from plasma samples and analysed by nanoflow ultra\u2010high\u2010performance liquid chromatography\u2013high\u2010resolution tandem mass spectrometry (nano\u2010UHPLC\u2010HRMSMS) using data\u2010independent acquisition. Differential protein abundance analysis has shortlisted seven protein biomarker candidates that showed significant changes specifically after Mircera administration in the treated but not in the control geldings, which comprised downregulation of two proteins, haptoglobin (HP) and haemopexin (HPX), and upregulation of five proteins, transferrin receptor 1 (TFR1), phospholipid transfer protein (PLTP), tenascin C (TNC), vascular cell adhesion molecule 1 (VCAM1) and galectin 3 binding protein (LGALS3BP). Multivariate analysis of plasma proteome has allowed the classification of control and treated samples. This is the first report on the discovery of plasma protein biomarkers of rhEPO administration to geldings. The results lay a foundation for applications of protein biomarkers for controlling the misuse of ESAs.<\/jats:p>","DOI":"10.1002\/dta.3810","type":"journal-article","created":{"date-parts":[[2024,9,22]],"date-time":"2024-09-22T20:24:21Z","timestamp":1727036661000},"page":"1053-1062","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Discovery of Biomarkers of a Recombinant Human Erythropoietin Administration to Thoroughbred Geldings by Label\u2010Free Proteomics"],"prefix":"10.1002","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2471-1790","authenticated-orcid":false,"given":"Hiu Wing","family":"Cheung","sequence":"first","affiliation":[{"name":"Racing Laboratory, The Hong Kong Jockey Club Sha Tin Racecourse  Sha Tin Hong Kong, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7136-4712","authenticated-orcid":false,"given":"Kin\u2010Sing","family":"Wong","sequence":"additional","affiliation":[{"name":"Racing Laboratory, The Hong Kong Jockey Club Sha Tin Racecourse  Sha Tin Hong Kong, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jimmy C. 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