Studies on the Gelation Reaction of Limulus Lysate (Pre-gel). V. Purification of the Inhibitory Substances of the Gelation Reaction of Limulus Lysate from Human Plasma

Publisher Website
Google Scholar
Abstract
The inhibitory substances of the gelation reaction of Limulus lysate [used in endotoxin assays] were purified from human plasma by a 5-step procedure. Two purified fractions, E-1 and E-3, were obtained and the recovery of the inhibitory activities in E-1 and E-3 was 8% and 2% respectively. When examined with disc gel electrophoresis and immunoelectrophoresis against anti-human plasma antiserum, E-3 migrated as a single component corresponding to .alpha.2-globulin and E-1 migrated as a major component corresponding to .alpha.1-globulin and minor components. The major electrophoretic component of E-1 represented 83% of the protein by gel scanning. The major component of E-1 gave a precipitin line against anti-human .alpha.1-antitrypsin antiserum and E-3 gave a precipitin line against anti-human antithrombin III antiserum by Ouchterlony immunodiffusion and immunoelectrophoresis. The results of MW, carbohydrate and amino acid analyses of E-1 and E-3 were similar to those of .alpha.1-antitrypsin and antithrombin III respectively. Purified .alpha.1-antitrypsin and antithrombin III inhibited the gelation reaction of Limulus lysate. The inhibitory substances of the gelation reaction of Limulus lysate in human plasma are .alpha.1-antitrypsin and antithrombin III and most of the inhibitory activity is attributable to .alpha.1-antitrypsin.

This publication has 5 references indexed in Scilit: