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. 1992 Jul;7(2):73-80.
doi: 10.3904/kjim.1992.7.2.73.

The effect of dilution, pH and ionic strength of plasma on t-PA precipitation in euglobulin fraction

S Y Hong  1 , H K Shin, D H Yang
Affiliations

The effect of dilution, pH and ionic strength of plasma on t-PA precipitation in euglobulin fraction

S Y Hong et al. Korean J Intern Med. 1992 Jul.

Abstract

In order to evaluate the influence of dilution, pH and ionic strength on the precipitation of t-PA and PAI-1 during euglobulin precipitation, we measured t-PA Ag, PAI-1 Ag and fibrinolytic activity in the euglobulin fraction made of pooled plasma from liver cirrhosis patients, under various conditions by changing pH, ionic strength and degree of dilution. The precipitation of t-PA Ag in the euglobulin fraction was enhanced by decreasing the ionic strength and greatest at pH 6.0. The fibrinolytic activity in the euglobulin fraction showed consistent changes with t-PA Ag under varying pH and ionic strength. The precipitation of t-PA Ag was not influenced by the dilution factor but the larger the dilution factor, the greater the PAI-1 and the smaller the fibrinolytic activity in the euglobulin fraction. PAI Ag in euglobulin fraction showed consistent changes with t-PA Ag in the euglobulin fraction regardless of the changes in ionic strength and pH. The amount of precipitation of t-PA and PAI-1 was increased by the presence of dextran sulfate, under varying pH, ionic strength and dilution conditions. Our results show that the currently used conditions for standard euglobulin precipitation are the most favorable for t-PA precipitation into the euglobulin fraction. The fibrinolytic activity exerted in the euglobulin fraction seems to depend on the amount of t-PA-PAI-1 complex rather than minimized protease inhibitor in the euglobulin fraction.

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Figures

Fig. 1.
Fig. 1.
The effect of ionic strength on the relationship between fibrinolytic activity and t-PA Ag in the euglobulin fraction. The dextran sulfate euglobulin fraction prepared at a constant pH (5.8–6.0) and 1:10 dilution. Variations in ionic strength is recorded on the abscissa as final concentration of added NaCl in 1:10 diluted plasma. t-PA Ag and fibrinolytic activity show a consistent change at low ionic concentration state.
Fig. 2.
Fig. 2.
The effect of pH on the relationship between fibrinolytic activity and t-PA Ag in euglobulin fraction. The dextran sulfate euglobulin fraction prepared at a constant ionic strength (0.15) and 1:10 dilution. Variation in pH is recorded on the abscissa. t-PA Ag and fibrinolytic activity sow a consistent change.
Fig. 3.
Fig. 3.
The effect of dilution on the relationship between fibrinolytic activity and t-PA Ag in euglobulin fraction. The dextran sulfate euglobulin fraction prepared at a constant ionic strength (0.15) and at pH 5.8–6.0. Variations in dilution is recorded on the abscissa. t-PA Ag show no significant change but the fibrinolytic activity decreases as the dilution factor increases.
Fig. 4.
Fig. 4.
The effect of ionic strength on the relationship between PAI-1 Ag and t-PA Ag in the euglobulin fraction with or without dextran sulfate. Euglobulin fraction prepared at a constant pH (5.8–6.0) and 10 times dilution with or without dextran sulfate. Variations in ionic strength is recorded on the abscissa as final concentration of added NaCl in 1:10 diluted plasma. t-PA Ag, PAI-1 Ag, with or without dextran sulfate show a consistent change at low ionic concentration state.
Fig. 5.
Fig. 5.
The effect of pH on the relationship between PAI-1 Ag and t-PA Ag in euglobulin fraction with or without dextran sulfate. Euglobulin fraction prepared at a constant ionic strength (0.15) and 1:10 dilution with or without dextran sulfate. Variation in pH is recorded on the abscissa. t-PA Ag, PAI-1 Ag, with or without dextran sulfate, show a consistent change.
Fig. 6.
Fig. 6.
The effect of dilution on the relationship between PAI-1 Ag and t-PA Ag in euglobulin fraction with or without dextran sulfate. Euglobulin fraction prepared at a constant ionic strength (0.15) and pH 5.8–6.0, with or without dextran sulfate. Variation in dilution is recorded on the abscissa. t-PA Ag shows no significant change but the PAI-1 Ag increases as the dilution factor increases.
Fig. 7.
Fig. 7.
The effect of ionic strength on the euglobulin fibrinolytic activity, with or witout dextran sulfate. The euglobulin fraction prepared at a constant pH (5.8–6.0). Variation in ionic strength is recorded on the abscissa as final concentration of added NaCl in 1:10 diluted plasma. The fibrinolytic activity is higher in dextran sulfate euglobulin fraction regardless of the change in ionic strength.
Fig. 8.
Fig. 8.
The effect of pH on the euglobulin fibrinolytic activity, with or without dextran sulfate. The euglobulin fraction prepared at a constant ionic strength (0.15) and 1:10 dilution. The fibrinolytic activity is higher in dextran sulfate euglobulin fraction at a pH between 5.0–9.0.
Fig. 9.
Fig. 9.
The effect of dilution on the euglobulin fibrinolytic activity, with or without dextran sulfate. The euglobulin fraction prepared at a constant ionic strengt (0.15) and pH 5.8–6.0. The fibrinolytic activity is higher in dextran sulfate euglobulin fraction regardless of the change of dilution ratio.
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