Two-Dimensional Differential Gel Electrophoresis to Identify Protein Biomarkers in Amniotic Fluid of Edwards Syndrome (Trisomy 18) Pregnancies

Abstract

BackgroundEdwards syndrome (ES) is a severe chromosomal abnormality with a prevalence of about 0.8 in 10,000 infants born alive. The aims of this study were to identify candidate proteins associated with ES pregnancies from amniotic fluid supernatant (AFS) using proteomics, and to explore the role of biological networks in the pathophysiology of ES.MethodsAFS from six second trimester pregnancies with ES fetuses and six normal cases were included in this study. Fluorescence-based two-dimensional difference gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) were used for comparative proteomic analysis. The identified proteins were further validated by Western blotting and the role of biological networks was analyzed.ResultsTwelve protein spots were differentially expressed by more than 1.5-fold in the AFS of the ES pregnancies. MALDI-TOF/MS identified one up-regulated protein: apolipoprotein A1 (ApoA1), and four under-regulated proteins: vitamin D binding protein (VDBP), alpha-1-antitrypsin (A1AT), insulin-like growth factor-binding protein 1 (IGFBP-1), and transthyretin (TTR). Western blot and densitometric analysis of ApoA1, A1AT, IGFBP-1, and TTR confirmed the alteration of these proteins in the amniotic fluid samples. Biological network analysis revealed that the proteins of the ES AFS were involved mainly in lipid and hormone metabolism, immune response, and cardiovascular disease.ConclusionsThese five proteins may be involved in the pathogenesis of ES. Further studies are needed to explore.

Vitamin D-binding protein in cervicovaginal fluid as a non-invasive predictor of intra-amniotic infection and impending preterm delivery in women with preterm labor or preterm premature rupture of membranes

“…In accordance with the results of the current study, the previous longitudinal study by Liong et al showed that VDBP is detectable in the CVF of all pregnant women during the second half of pregnancy, with no change in the CVF VDBP level between 20 and 35 weeks’ gestation [ 15 ], suggesting that VDBP is a physiologic constituent of the lower genital tract fluid. In addition, VDBP has been previously identified in the AF of pregnant women [ 19 ], and our preliminary experiment to determine the dilution ratio showed that VDBP levels in the AF without intra-amniotic infection were approximately 10 times higher than the levels in the CVF (supplementary materials). Similarly, the VDBP levels reported in pregnant maternal serum were 1000 times higher than those measured in the CVF in the current study [ 20 , 21 ].…”

Vitamin D-Binding Protein in Cervicovaginal Fluid as a Non-Invasive Predictor of Maternal and Fetal Outcome in Women with Preterm Labor

“…In accordance with the results of the current study, the previous longitudinal study by Liong et al showed that VDBP is detectable in the CVF of all pregnant women during the second half of pregnancy with no change in the CVF VDBP level between 20 and 35 weeks' gestation [14] suggesting that VDBP is a physiologic constituent of the lower genital tract fluid. In addition, VDBP has been previously identified in the AF of pregnant women [8] and on other experiment to determine the dilution ratio showed that VDBP levels in the AF without intra-amniotic infection were approximately 10 times higher than the levels in the CVF (supplementary materials) [12] . Similarly, the VDBP levels reported in pregnant maternal serum were 1000 times higher than those measured in the CVF in the current study [3] .…”

Preliminary study of protein changes in trisomy 21 fetus by proteomics analysis in amniocyte

“…MALDI‐TOF MS analysis was performed as a previous study . Pancreatin was used to calibrate ABI 4800M MALDI MS analyzer equipped with a 200 Hz 366 nm N 2 laser.…”

“…MALDI-TOF MS analysis was performed as a previous study. 8 Pancreatin was used to calibrate ABI 4800M MALDI MS analyzer equipped with a 200 Hz 366 nm N 2 laser. The MALDI-TOF MS test was performed at a mass range of 700 to 3200 Da (best mass range at 1500 Da).…”