One of the most urgent requirements in prostate cancer diagnosis is the development of a blood-based test which would be able to distinguish prostate cancer from benign prostate hyperplasia. Previously published results found a significant difference between specific glycan levels in patients with advanced prostate cancer and healthy controls. N-glycans from the whole serum glycoproteins were measured using our fully quantitative high throughput N-glycan analysis in combination with exoglycosidase digestions in sera from 13 benign prostate hyperplasia and 34 prostate cancer samples (17 Gleason score 5 and 17 Gleason score 7). The levels of core fucosylated biantennary glycans and alpha2-3 linked sialic acids were significantly increased in prostate cancer patients compared to patients with benign prostate hyperplasia. Triantennary trigalactosylated glycans and tetraantennary tetrasialylated glycans with outer arm fucose were significantly decreased and tetraantennary tetrasialylated glycans increased in Gleason 7 compared to Gleason 5. All these glycans can distinguish prostate cancer patients from benign prostate hyperplasia or Gleason 7 from Gleason 5 prostate cancer patients better than the current clinical test prostate-specific antigen, therefore their measurement may provide a new non-invasive approach to diagnose prostate cancer. However, additional validation studies would need to be carried out to further support this finding. Decreases in triantennary trigalactosylated glycans and/or bisected core fucosylated biantennary monosialylated glycans and increases in tetraantennary tetrasialylated glycans correlate with perineural invasion which could further help to diagnose tumour spread and predict patients' survival.One of the most urgent requirements in prostate cancer diagnosis is the development of a blood-based test which would be able to distinguish prostate cancer from benign prostate hyperplasia. Previously published results found a significant difference between specific glycan levels in patients with advanced prostate cancer and healthy controls. N-glycans from the whole serum glycoproteins were measured using our fully quantitative high throughput N-glycan analysis in combination with exoglycosidase digestions in sera from 13 benign prostate hyperplasia and 34 prostate cancer samples (17 Gleason score 5 and 17 Gleason score 7). The levels of core fucosylated biantennary glycans and alpha2-3 linked sialic acids were significantly increased in prostate cancer patients compared to patients with benign prostate hyperplasia. Triantennary trigalactosylated glycans and tetraantennary tetrasialylated glycans with outer arm fucose were significantly decreased and tetraantennary tetrasialylated glycans increased in Gleason 7 compared to Gleason 5. All these glycans can distinguish prostate cancer patients from benign prostate hyperplasia or Gleason 7 from Gleason 5 prostate cancer patients better than the current clinical test prostate-specific antigen, therefore their measurement may provide a new non-invasive approach to diagnose prostate cancer. However, additional validation studies would need to be carried out to further support this finding. Decreases in triantennary trigalactosylated glycans and/or bisected core fucosylated biantennary monosialylated glycans and increases in tetraantennary tetrasialylated glycans correlate with perineural invasion which could further help to diagnose tumour spread and predict patients' survival.