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Contributions to the Study of Plasminogen: High Purification of Human Plasminogen
Author(s) -
Stoian Florica Ionescu,
Istrati Maria
Publication year - 1968
Publication title -
vox sanguinis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.68
H-Index - 83
eISSN - 1423-0410
pISSN - 0042-9007
DOI - 10.1111/j.1423-0410.1968.tb01735.x
Subject(s) - chromatography , chemistry , column chromatography , sephadex , elution , agarose , acetic acid , polyacrylamide , electrophoresis , phosphate buffered saline , hydrolysis , phosphate , biochemistry , enzyme , polymer chemistry
Summary. By means of series chromatography on Sephadex G‐100 gel columns the authors have prepared highly purified human plasminogen from human plasma. Initially, the plasma proteins are fractionated on Sephadex G‐100 columns utilizing 0.002 M alkaline arginine solution pH 9 as eluent; enzyme‐active effluents are precipitated at pH 6 with 1% solution of acetic acid; partly purified plasminogen precipitate is dissolved in 0.15 M phosphate buffer, pH 7.6. The plasminogen solution is further submitted to chromatography on two series of Sephadex G‐100 columns, using 0.15 M saline phosphate buffer solution, pH 7.6, as eluent. The highly purified plasminogen resulting from the effluents which form the descending slope of the elution curves on the last column series is lyophilized, and has a specific activity of 18 cas. U/mg of protein. The lyophilized preparation is immunoelectrophoretically and electrophoretically homogeneous; acrylamide‐agarose mixed gel electrophoresis shows a single band. Tiselius free electrophoresis of the plasminogen 0.8% solution in saline phosphate buffer pH 7.6 shows a single sharp peak. The plasminogen preparation is soluble at neutral pH, and at acid pH in saline buffer solutions. The preparation possesses the required characteristics for structural studies.