Chromatographic isolation and determination of the concentration in human plasma of the novel plasma protein, sgp120

Published

Journal Article

sgp120 is a recently discovered normal human plasma protein. The physiological Function of this abundant protein is still unresolved but supposedly lies at the borderline of the coagulation contact system and the complement system. In this paper the combined chemical-chromatographics method for the isolation and purification of sgp120 is presented. sgp120 was isolated from the stabilized normal human plasma by two subsequent precipitations with ammonium sulphate (1.2 and 2.2 mol/L), removal of the euglobulins, fractionation over an anion exchange column (DEAE-Sephacel), Jacalin-Agarose affinity chromatography and the gelfiltration over Sephacryl S-200. EPLC system (Pharmacia) was used for chromatographic procedures. The goat was immunized with the purified protein and the polyclonal antiserum thus obtained was used for the determination of sgp120 concentrations in plasma samples. Two methods were used for this purpose (radial immunodiffusion and Laurell's 'rocket' electrophoresis) giving the same results. Higher than normal concentrations were detected in the cases of sgp120 degradation during the contact system activation. Thus, the use of Western blot was necessary to follow sgp120 degradation pattern during the determination of sgp120 plasma concentration.

Cited Authors

  • Miletic, VD; Frank, MM

Published Date

  • December 1, 1994

Published In

  • Jugoslovenska Medicinska Biokemija

Volume / Issue

  • 13 / 3-4

Start / End Page

  • 95 - 101

International Standard Serial Number (ISSN)

  • 0352-1311

Citation Source

  • Scopus