Abstract
Early detection of any preeclampsia biomarkers may lower the risk of mortality, both for a mother and a child. Our study focuses on techniques for preeclampsia biomarker identification by comparing the results of a method using liquid chromatography mass spectrometry in multiple reaction monitoring mode (LC-MS/MS) with those by the enzyme-linked immunosorbent assay (ELISA) test, as well as by comparing the obtained results with clinical data. In the proposed LC-MS/MS method a tryptic digest peptide charge derivatization strategy was used as a tool for sensitive detection of podocin, i.e., a previously discovered preeclampsia biomarker present in urine samples from pregnant women. Urine samples from pregnant women with diagnosed preeclampsia were collected at different stages of pregnancy and from healthy subjects, and then were analyzed by ELISA test and the proposed method with LC-MS/MS. Charge derivatization of the ε amino group of C-terminal lysine residues in tryptic digests by 2,4,6-triphenylpyrylium salt was performed to increase the ionization efficiency in the LC-MS/MS mode. Podocin was identified at the early stage of pregnancy, while its detection using an ELISA test was not possible. The protocol for urine sample preparation was optimized. Our results show that the proposed method by LC-MS/MS in combination with peptide charge derivatization, provides an ultrasensitive tool for diagnosis of preeclampsia, and provides earlier detection than a clinical diagnosis or ELISA test. The proposed solution may revolutionize medical diagnostics.
Highlights
Prevention is better than the cure; prediction of disease or a fast and accurate diagnosis is crucial for selecting the most effective therapy
In the last two years, we developed an LC-Mass spectrometry (MS)/MS method for the identification of podocin tryptic peptide in feline [23], canine [24], and equine [25] urine samples, using urine sediment samples obtained from animals with diagnosed chronic kidney disease (CKD) and acute kidney injury (AKI)
We explored the applicability of a proposed method that combined peptide charge derivatization and Liquid Chromatography Mass Spectrometry (LC-MS)/MS as compared with the enzyme-linked immunosorbent assay (ELISA) test for early detection of PE in urine sediment samples from pregnant woman
Summary
Prevention is better than the cure; prediction of disease or a fast and accurate diagnosis is crucial for selecting the most effective therapy. New specific biomarkers are urgently needed to improve diagnostic methods; methods for the development and selection of reliable biomarkers are still challenging. They have been increasing steadily over the past decade [1]. The detection of podocytes in urine is a promising diagnostic test suitable for early detection of kidney damage [2]. Numerous reports have indicated that early diagnosis of elevated urinary podocyte excretion can predict an acute phase of kidney injury and may facilitate the diagnosis of kidney disease in humans as well as animals [9]. Results can be compared despite differences in the urine concentration, which is significant in the case of patients with congestive heart failure (CHF) and chronic kidney disease (CKD), who have disturbances of urine dilution due to the disease itself or prescribed medication
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