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KMID : 0624219980060000032
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Focus on Genetic Science
1998 Volume.6 No. 0 p.32 ~ p.39
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Reconstitution of Biologically Active FⅧ From Its Subunits
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Song, Dong-Weon
Lee, Mi-Young/Oh, Sang-Hwan
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Abstract
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Factor Ⅷ (FⅧ), a plasma glycoprotein involved in the process of blood coagulation cascade is composed of two subunits, heavy and light chains (FⅧ-H, FⅧ-L). Genetic defect in FⅧ gene is responsible for hemophilia A that may be alleviated by the supplementation of plasma derived FⅧ concentrate. Recombinant FⅧ (r FⅧ) produced in mammalian cell expression systems has been known to be useful for the substitution of plasma-derived FⅧ but some problems associated with clinical use remain yet. a biologically active FⅧ was reconstituted from affinity column-purified FⅧ subunits (FⅧ-H & FⅧ-L) and from rFⅧ-H and rFⅧ-L in the presence of CaCl₂(40mM) with FVIIII-depleted human plasma. the reconstituted plasma exhibited cofactor activity of FⅧ(0.7 units/㎖) and the in vitro bilogical half-life of the reconstituted FⅧ was relatively longer than that of wild type FⅧ. These results implicate that reconstituted FⅧ from its recombinant subunits (rFⅧ-H, rFⅧ-L) produced in the cell culture system may be an useful candidate for the treatment of hemophilia A.
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