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Label-free Proteomics for Discovering Biomarker Candidates for Controlling Krypton Misuse in Castrated Horses (Geldings)
journal contribution
posted on 2020-02-25, 20:04 authored by Kin-Sing Wong, Hiu Wing Cheung, Timmy L. S. Choi, Wai Him Kwok, Peter Curl, Stewart C. Mechie, Anil Prabhu, Terence S. M. Wan, Emmie N. M. HoRecent
advances in label-free quantitative proteomics may support
its application in identifying and monitoring biomarkers for the purpose
of doping control in equine sports. In this study, we developed a
workflow of label-free quantitative proteomics to propose plasma protein
biomarkers in horses after administration with krypton (Kr), a potential
erythropoiesis-stimulating agent. Plasma proteomes were profiled by
using nanoliquid chromatography–high-resolution mass spectrometry.
An in-house mass spectral library consisting of 1121 proteins was
compiled using samples collected from geldings (castrated horses)
in the administration trial and geldings in training. A data-independent
acquisition method was used to quantify an array of plasma proteins
across plasma samples from the administration trial. Statistical analyses
proposed a profile of 83 biomarker candidates that successfully differentiated
Kr-administered samples from control samples, with the ability to
detect Kr exposure for up to 13 days (the last sample collected in
the administration trial). The model also correctly classified 32
in-training geldings as untreated controls. This is significantly
longer than the 1 h detection time of plasma Kr using headspace gas
chromatography–tandem mass spectrometry. Bioinformatic analyses
enriched biomarker candidates relevant to complement activation and
iron metabolism. The upregulation of transferrin receptor protein
1, one of the candidates related to iron metabolism, in plasma after
Kr administration was validated by selected reaction monitoring of
corresponding peptides. These results have demonstrated label-free
quantitative proteomics as a promising approach to propose plasma
protein biomarkers to enhance doping control. Data are available via
ProteomeXchange with identifier PXD017262.