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by Alan E. Mast, Jason E. Stadanlick, J. Marcus Lockett, and Dennis J

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1 Solvent/Detergent-Treated Plasma Has Decreased Antitrypsin Activity and Absent Antiplasmin Activity
by Alan E. Mast, Jason E. Stadanlick, J. Marcus Lockett, and Dennis J. Dietzen Blood Volume 94(11): December 1, 1999 ©1999 by American Society of Hematology

2 Noncovalent polymers of antitrypsin and, to a much lesser extent, antithrombin, are present in S/D plasma. Noncovalent polymers of antitrypsin and, to a much lesser extent, antithrombin, are present in S/D plasma. Western blot analysis of plasma proteins. (A) Nondenaturing-PAGE and (B) SDS-PAGE with molecular weight standards as indicated: (1) antithrombin in FFP; (2) antithrombin in S/D plasma; (3) antitrypsin in FFP; (4) antitrypsin in S/D plasma. Alan E. Mast et al. Blood 1999;94: ©1999 by American Society of Hematology

3 The latent and polymerized conformations of antiplasmin are present in S/D plasma.
The latent and polymerized conformations of antiplasmin are present in S/D plasma. Western blot analysis of plasma proteins. (A) Nondenaturing-PAGE: (1) antiplasmin in FFP; (2) antiplasmin in FFP following S/D treatment in our laboratory (see Materials and Methods); (3) antiplasmin in S/D plasma; (4) antiplasmin in FFP diluted 10-fold more than lane 1. (B) SDS-PAGE with molecular weight standards as indicated: (1) antiplasmin in FFP; (2) antiplasmin in S/D plasma. (C) Nondenaturing-PAGE of S/D plasma containing aggregates: (1) antiplasmin in S/D plasma after removal of aggregates; (2) antiplasmin in aggregates isolated as described in Materials and Methods. Alan E. Mast et al. Blood 1999;94: ©1999 by American Society of Hematology

4 Conformational stability of antithrombin, antitrypsin, and antiplasmin in FFP and S/D plasma.
Conformational stability of antithrombin, antitrypsin, and antiplasmin in FFP and S/D plasma. Analysis is performed by TUG-PAGE. Each panel represents Western blot analysis of plasma proteins separated on a single gel with the direction of migration from top to bottom and the urea gradient (0 to 8 mol/L) from left to right. Alan E. Mast et al. Blood 1999;94: ©1999 by American Society of Hematology

5 Progress curve amidolytic assays measuring inhibition of PPE or plasmin by FFP and S/D plasma.
Progress curve amidolytic assays measuring inhibition of PPE or plasmin by FFP and S/D plasma. Reactions were initiated by the addition of either 0.4 μmol/L PPE or 0.2 μmol/L plasmin to a mixture of plasma and 500 μmol/L of the appropriate substrate. (A) Reactions initiated with PPE: (1) noninhibited control; (2) S/D plasma; (3) 1:2 dilution of FFP; (4) FFP; (5) S/D plasma after addition of 12.5 μmol/L purified antitrypsin. (B) Reactions initiated with plasmin: (1) noninhibited control; (2) S/D plasma; (3) 1:10 dilution of FFP; (4) FFP; (5) S/D plasma after addition of 0.9 μmol/L purified antiplasmin. Alan E. Mast et al. Blood 1999;94: ©1999 by American Society of Hematology


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