Two Fractions of Macroglobulin XaI isolated by CM-cellulose Chromotography

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Human citrated plasma was treated with ammonium sulfate at 30-50% saturation, and applied to the column of sephadex G-200. Macroglobulin XaI, which was obtained by gel filtration on Sephadex G-200, was separated to two fractions by CM-cellulose chromatography. One fraction (Fr. A) was eluted with pH 6.0, 0.02M acetate buffer, and another fraction (Fr. B) with 0.06M NaCl in the same buffer by 0→0.2M NaCl gradient elution on CM-cellulose. Rechromatography of Fr. A and Fr. B resulted in the same elution pattern as the first chromatography, namely it was obtained Fr. A and Fr. B from Fr. A, and Fr. B (a) and Fr. B (b) from Fr. B.These observations have given rise to the speculation that anti-Xa substance in Fr. A may be identical with Fr. B.Fr. A showed one main band and many other bands on polyacrylamide gel disc electrophoresis, and three precipitin lines besides α2-macroglobulin on immunoelectrophoresis. While, Fr. B (Fr. B (a), Fr. B (b)) with anti-Xa activity showed one main band and two other thin bands on polyacrylamide gel disc electrophoresis, but only one distinct precipitin line (α2-macroglobulin) on immunoelectrophoresis. These observations indicate that anti-Xa substance in macroglobulin fraction is α2-macroglobulin.
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