To identify biochemical markers in men with idiopathic infertility and normal sperm counts. We obtained proteomic profiling proteins in human spermatozoa following successful or unsuccessful pregnancy via assisted reproductive technology (ART) using 6-plex tandem mass tag (TMT) isobaric mass spectrometry. Our study design consisted of two groups: 1. The semen of 6 men whose sperm resulted in a clinical pregnancy following ART and 6 men whose semen did not result in a clinical pregnancy following ART. The results of differentiated mass spectrometry were validated by Western blotting. A total of 2,045 proteins were detected in our cohort. 21 proteins were found to be differentially expressed (>1.2-fold) in men whose sperm resulted in a clinical pregnancy and those that did not. Using the results of bioinformatics analysis and Western Blotting, three proteins (A2LD1, ATP1B3 and FBXO2) were shown to have the same differential pattern (p<0.05) that was observed in the mass spectrometry analysis. Proteomics may help identity a select cohort of men with abnormal semen parameters and aide infertility diagnoses.
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