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KMID : 0391319940040020301
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Korean Journal of Biological Response Modifiers
1994 Volume.4 No. 2 p.301 ~ p.306
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Receptor Binding of DA-3030, A Recombinant Human Granulocyte Colony-stimulating Factor, to Normal Murine Bone Marrow Cells
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손미원
강수형/김원배/양중익
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Abstract
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Background:
@EN Human granulocyte colony0stimulating factor (G-CSF) is a regulatory glycoprotein that stimulates the production of neutrophilic granulocytes from committed hematopoietic progenitor cells and G-CSF, like each of the other CSFs and ILS, exerts
its
biologic activities through binding to CSF-specific high affinity receptors. In this report, the binding of DA-3030, a recombinant human G-CSF, to murine bone marrow cells was investigated using a radioiodinated DA-3030 with high speific
radioactivity.
@ES Methods:
@EN DA-3030 was radioiodinated using iodobead and carrier-free Na125 I. Bone marrow cells were obtained from femurs of mice. The binding experiments of 125I-labelled G-CSF to murine bone marrow cells was performed.
@ES Results:
@EN The radioiodinated preparation of DA-3030 showed high specific activity (50,000cpm/ng) and retained full biological activity for at least 3 days. The binding was time and temperature-dependent, saturable and highly specific. Unlabeled DA-3030
and
filgrastim equally competeted for the binding of labeled DA-3030 in dose-dependent manner, demonstrating that DA-3030 binded to target cells, as like as filgrastim.
Scatchard analysis of the specific bindings of labeled ligands revealed a single class of binding site with an apparent dissociation constant (kd) of 50~60pM and 100~200 maximal binding sites per cell.
@ES Conclusion:
@EN These data indicate that the radioiodinated preparation of DA-3030 binds to the same specific receptor with same affinity as filgrastim or human G-CSF. These results suggests that DA-3030 is suitable for studies on human G-CSF receptors of
various
cells and provide more useful information for clinical application of rhG-CSF.
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KEYWORD
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