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Blood, Vol. 95 No. 9 (May 1), 2000:
pp. 2975-2982
The assessment of serum nontransferrin-bound iron in chelation
therapy and iron supplementation
William Breuer,
Aharon Ronson,
Itzchak N. Slotki,
Ayala Abramov,
Chaim Hershko, and
Z.
Ioav Cabantchik
Department of Biological Chemistry, Institute of Life Sciences,
Hebrew University of Jerusalem, Jerusalem, Israel; Department of
Internal Medicine and Nephrology Unit, Department of Pediatrics, Shaare
Zedek Medical Center, Jerusalem, Israel.
Nontransferrin-bound iron (NTBI) appears in the serum of individuals
with iron overload and in a variety of other pathologic conditions.
Because NTBI constitutes a labile form of iron, it might underlie some
of the biologic damage associated with iron overload. We have developed
a simple method for NTBI determination, which operates in a 96-well
enzyme-linked immunosorbent assay format with sensitivity comparable to
that of previous assays. A weak ligand, oxalic acid, mobilizes the NTBI
and mediates its transfer to the iron chelator deferoxamine (DFO)
immobilized on the plate. The amount of DFO-bound iron, originating
from NTBI, is quantitatively revealed in a fluorescence plate reader by
the fluorescent metallosensor calcein. No NTBI is found in normal sera
because transferrin-bound iron is not detected in the assay. Thalassemic sera contained NTBI in 80% of the cases (range, 0.9-12.8 µmol/L). In patients given intravenous infusions of DFO, NTBI initially became undetectable due to the presence of DFO in the sera,
but reappeared in 55% of the cases within an hour of cessation of the
DFO infusion. This apparent rebound was attributable to the loss of DFO
from the circulation and the possibility that a major portion of NTBI
was not mobilized by DFO. NTBI was also found in patients with
end-stage renal disease who were treated for anemia with intravenous
iron supplements and in patients with hereditary hemochromatosis, at
respective frequencies of 22% and 69%. The availability of a simple
assay for monitoring NTBI could provide a useful index of iron status
during chelation and supplementation treatments.
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