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Recent data indicated that H-FABP is a promising plasma biomarker for early and sensitive diagnosis of acute coronary heart syndrome and myocardial infarction in heart failure (33) and that B-FABP may be a plasma biomarker for the detection of brain injury (36).
7] Nonstandard abbreviations: A-, H-, L-, E-, I-, and B-FABP, adipocyte, heart-type, liver-type, keratinocyte, intestine-type, and brain-type fatty acid-binding protein, respectively; apo E, apolipoprotein E; 2-DE, 2-dimensional gel electrophoresis; MS/MS, tandem mass spectrometry; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; BMI, body mass index; and HOMA, homeostasis model assessment.
B-FABP and H-FABP are members of a family of nine distinct FABP types, each named after the tissue in which it was first detected (16).
The aim of our study was to investigate the tissue distribution and concentrations of B-FABP and H-FABP in human brain and to study the potential of these proteins as serum markers for the detection of brain injury in patients.
To determine the biological variation for circulating B-FABP and to establish a reference interval, we obtained plasma samples (EDTA) from 80 healthy individuals (40 males and 40 females; age ranges, 21-30, 31-40, 41-50, 51-60, and 61-70 years; n = 8 for each group) visiting the blood bank of Litige to study the influence of age and gender, and from another 12 healthy individuals (citrated plasma; 6 males and 6 females; age range, 19-27 years) from the Maastricht University student population to study the influence of circadian rhythm (21).
Production of recombinant B-FABP and antibodies against B-FABP.
5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, blotted the gels on a HyBond nitrocellulose membrane (Amersham Biosciences), and detected the FABPs with purified polyclonal antibodies against B-FABP and with monoclonal antibodies against H-FABP (67D3 and 66E2 as capture and detector antibodies, respectively, in the H-FABP ELISA; HyCult Biotechnology) (23,24).
After the wells were washed with PBS, the B-FABP calibrator and two- to fivefold-diluted samples in sample buffer (PBS containing 5 g/L bovine serum albumin and 0.
Western blot analysis showed that the monoclonal anti-H-FABP antibody 67D3, used as capture antibody in the ELISA, cross-reacts slightly with B-FABP (Fig.
In each part of the human brain studied, the tissue concentration of H-FABP was found to be >10-fold higher than that of B-FABP (Fig.
Plasma B-FABP concentrations in healthy individuals were <5 [micro]g/L, the detection limit of the assay.
B-FABP serum concentrations were increased (>5 [micro]g/L) in 88 of 130 patients with MTBI, whereas H-FABP (>6 [micro]g/L) was increased in 91 patients (Table 1).
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