Urinary Proteomic/Peptidomic Biosignature of Breast Cancer Patients Using 1D SDS-PAGE Combined with Matrix-Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry

Abstract

The potential development of a rapid and highly sensitive breast cancer (BC) diagnostic method has been increasingly investigated by many researchers in order to significantly improve the diagnosis of this disease that affects millions of women worldwide. Thus, this investigation aimed to establish a potential BC urinary peptidomic pattern using one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (1D SDS-PAGE) coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) as a useful approach for BC diagnosis. The results of Lowry’s assay demonstrated that the total protein concentration increased after precipitation and that the healthy control group (HCs, 160 to 318 µg/mL, 142 ± DD µg/mL, on average) presented higher total protein content than the BC patients (140 to 311 µg/mL, 115 ± DD µg/mL, on average). Related to MALDI-TOF MS analysis, the results revealed that four peptide ion biosignatures (m/z 1046.5, 1062.5, 1237.7 and 1727.9) allowed the discrimination between BC patients and HCs. The distinction efficiency and accuracy of BC urine peptides were determined by receiver operating characteristic (ROC) curve analysis that enabled the recognition of some features with great sensitivity (88%) and specificity (98%). Therefore, the obtained data revealed MALDI-TOF MS as a powerful tool to explore peptidomic biosignatures due to its speed, sensitivity, and mass accuracy, which allow the establishment of novel disease biomarkers.

Recent Contributions of Mass Spectrometry-Based “Omics” in the Studies of Breast Cancer

“…Four peptide ion biosignatures were identified that can discriminate between BC patients and controls. 180 Nipple aspirate fluid (NAF) analysis can be a noninvasive method for BC diagnosis. In a paired-proteomic shotgun strategy applied on NAF collected from breasts of patients, 1227 proteins were identified and quantified.…”

“…Noninvasive diagnosis and identification based on differential proteomes of biofluids have been attempted in various studies. Such elucidations are from tears, saliva, urine; ,, VOC of breath samples and urine samples; and nipple aspirates . The breast milk proteome signature from BC patients may be used as a diagnostic identifier of lactating women .…”

“…The total protein concentration was found to be high in normal control groups (160 to 318 μg/mL) compared to that in BC patients (140 to 311 μg/mL). Four peptide ion biosignatures were identified that can discriminate between BC patients and controls . Nipple aspirate fluid (NAF) analysis can be a noninvasive method for BC diagnosis.…”

MALDI-TOF MS: A Promising Analytical Approach to Cancer Diagnostics and Monitoring

“…MALDI-TOF MS has the potential to revolutionize cancer diagnostics by enabling biomarker discovery, enabling tissue imaging, and quantifying biomarker levels. However, the use of MALDI-TOF MS as a diagnostic tool for cancer is still evolving, and there are some critical assessments that need to be addressed, including further validation, standardization, data interpretation, optimization of sensitivity and specificity, cost considerations, and expanding biomarker coverage [36,136]. In this sense, the standardization of sample preparation, instrument calibration, and data analysis protocols is crucial to ensure the reproducibility and reliability of the data obtained [136,137].…”