Discovery and validation of new protein biomarkers for urothelial cancer: a prospective analysis

A Distinct Urinary Biomarker Pattern Characteristic of Female Fabry Patients That Mirrors Response to Enzyme Replacement Therapy

“…In terms of diagnostic accuracy, CE-MS also favourably compares to urinary Gb 3 , for which an AUC of 0.876 has been reported [33] (vs. 0.970 for CE-MS in our study), although lyso-Gb 3 seems to perform better than total Gb 3 [34]. In addition, because similarly accurate CE-MS based diagnostic models have been developed for a variety of other renal and cardiovascular diseases [13], [35], [36], [37], [38], [39], CE-MS may give hints to an alternative diagnosis in Fabry negative patients, i.e. this approach may offer an efficient diagnostic method, which can detect a variety of diseases using a single diagnostic test.…”

“…Control urine samples were taken from healthy volunteers and patients with a variety of other renal, metabolic and cardiovascular diseases. These samples have been previously collected as part of several clinical studies (refs [13], [35], [36], [37], [38], [39] and as yet unpublished studies). All samples were collected in the morning from midstream urine, frozen at −20°C without prior centrifugation and without addition of protease inhibitors, as suggested in recently published recommendations [42].…”

“…CE-MS analysis was performed as described previously [13], [43] using a P/ACE MDQ capillary electrophoresis system (Beckman Coulter, Fullerton, USA) on-line coupled to a Micro-TOF MS (Bruker Daltonic, Bremen, Germany). The ESI sprayer (Agilent Technologies, Palo Alto, CA, USA) was grounded, and the ion spray interface potential was set between −4 and −4.5 kV.…”

“…Spectra were accumulated every 3 s, over a range of m/z 350 to 3000. Accuracy, precision, selectivity, sensitivity, reproducibility, and stability of the analytical platform were demonstrated and described in great detail elsewhere [13].…”

“…Capillary electrophoresis coupled to mass spectrometry (CE-MS) is a reliable high-throughput method to simultaneously measure the excretion of hundreds of polypeptides and small proteins in the urine [13]. This technique utilizes capillary electrophoresis (CE) to separate small proteins according to their electrophoretic characteristics, directly followed by mass spectrometric analysis (MS) by electron spray ionization (ESI) and time-of-flight mass spectrometry (TOF-MS).…”