Two‐dimensional protein electrophoresis and multiple hypothesis testing to detect potential serum protein biomarkers in children with fetal alcohol syndrome

Abstract

Fetal alcohol syndrome (FAS) surveillance and intervention efforts are hampered by the lack of a specific biochemical test for diagnosis of the syndrome. Based on the hypothesis that abnormalities in growth and development (key features of FAS) involve altered protein metabolism, we analyzed serum proteins by two-dimensional gel electrophoresis and image analysis to search for potential protein biomarkers of FAS. Serum samples from 12 participants in whom FAS had been diagnosed and 8 sex- and age-matched participants whose mothers did not consume alcohol were analyzed in duplicate to determine whether the integrated intensities of matched proteins are significantly altered in children with FAS. Multiple hypothesis testing on 34 of the gels consisting of more than 1700 spots per gel revealed 21 proteins that we classified as potential protein biomarkers of FAS on the basis of significant t-test differences at p < 0.02. We classified 8 of the proteins as candidate biomarkers on the basis of significant concentration differences between case and control subjects at p < 0.01. One of the proteins is clearly an isoform of retinol binding protein; two appear in the area of the gel where alcohol dehydrogenase is expected to appear; one appears to be an isoform of alpha-1-antitrypsin; three appear to be isoforms of the beta-chain of haptoglobin; three may be forms of immunoglobulin light chains; and several others have not been associated with known proteins. No single protein differentiated all case subjects from control subjects, but stepwise canonical discriminant analyses revealed four groups of spots that distinguished between FAS case and control subjects with no misclassifications.(ABSTRACT TRUNCATED AT 250 WORDS)

Diagnostic Values of Surface‐Enhanced Laser Desorption/Ionization Technology for Screening of Habitual Drinkers

“…Two-dimensional electrophoresis(2-DE) has been the mainstay of proteomic studies and has been used to search for potential protein biomarkers for fetal alcohol syndrome (Robinson et al, 1995). Compared with 2-DE, SELDI-TOF MS combining chromatography and mass spectrometry is a more sensitive and rapid method to analyze complex mixtures of proteins and peptides.…”

2-D DIGE uterine endothelial proteomic profile for maternal chronic binge-like alcohol exposure

“…There is a great need for conducting proteomic studies to comprehend the mechanistic perspectives underlying the pathogenesis of FASD [15–19]. The current study is the first in the FASD research area to utilize quantitative 2-D DIGE methodology for this purpose.…”

“…It is evident that very few proteomic studies have been reported in the field of FASD research and they were conducted in an effort to discover biomarkers [15–19]. The earliest study in this field used two-dimensional gel electrophoresis and image analysis on serum samples obtained from FASD children and discovered that no single protein uniquely identified the condition [15].…”

“…The earliest study in this field used two-dimensional gel electrophoresis and image analysis on serum samples obtained from FASD children and discovered that no single protein uniquely identified the condition [15]. Datta and coworkers subsequently utilized MALDI-TOF/tandem MS and demonstrated decreases in amniotic fluid alpha fetoprotein using a FASD mouse model [16].…”

Application of proteomic technologies to discover and identify biomarkers for excessive alcohol consumption: A review

“…More recently, proteomic study was conducted again by 2-DE to search for potential protein biomarkers for fetal alcohol syndrome (FAS) [25]. Sera from 12 subjects in whom FAS had been diagnosed and 8 sex-and age-matched subjects whose mother did not consume alcohol were analyzed.…”