Subscribe to RSS
DOI: 10.1055/s-0038-1661096
Studies on the Procurement of Coagulation Factor VIII: Selective Precipitation of Factor VIII with Hydrophilic Polymers
Publication History
Received 05 March 1984
Accepted 23 March 1984
Publication Date:
19 July 2018 (online)
Summary
Early work on the purification of factor VIII using polyethylene glycol (PEG) indicated that other polymers might also be used to precipitate factor VIII leaving fibrinogen in solution. Recently polyvinylpyrrolidone (PVP) has also been advocated for this purpose.
In this study, different concentrations and molecular weights of hydroxyethyl starch, dextran, PEG, PVP, Ficoll, Percoll and albumin were examined for their ability to precipitate the factor VIII complex from cooled (not frozen) fresh CPD plasma. At optimal concentrations near quantitative recovery of VIIIR:Ag and VIII: CAg was achieved in 2 hr with minimal precipitation of fibrinogen or total protein. The best separations were observed with hydroxyethyl starch, albumin or Ficoll. PVP and PEG gave inferior purifications. Results for cryoprecipitate and intermediate-purity factor VIII concentrate were inferior to those obtained with plasma. Simple pre-concentration of plasma prior to chilling is an attractive alternative for large scale continuous production.
-
References
- 1 Pool JG, Shannon AE. Production of high-potency concentrates of antihaemophilic globulin in a closed-bag system; assay in vitro and in vivo. N Eng J Med 1965; 273: 1443-1447
- 2 Pool JG, Hershgold EJ, Pappenhagen AR. High-potency antihaemophilic factor concentrate prepared from cryoglobulin precipitate. Nature 1964; 203: 312
- 3 Newman J, Johnson AJ, Karpatkin M, Puszkin S. Methods for the production of clinically effective intermediate and high purity factor VIII concentrate. Br J Haematol 1971; 21: 1-20
- 4 Mason EC. Thaw-siphon technique for production of cryoprecipitate concentrate of factor VIII. Lancet 1978; 2: 15-19
- 5 Prowse CV, McGill A. Evaluation of the ‘Mason’ (continuous-thawsiphon) method for cryoprecipitate production. Vox Sang 1979; 37: 235-243
- 6 Rock GA, Cruikshank WH, Tackaberry ES, Palmer DS. Improved yields of factor VIII from heparinized plasma. Vox Sang 1979; 36: 294-300
- 7 Polson A, Potgieter GM, Largier JF, Mears GE F, Joubert FJ. The fractionation of protein mixtures by linear polymers of high molecular weight. Biochim Biophys Acta 1964; 82: 463-475
- 8 Hao YC, Ingham KC, Wickerhauser M. Fractional precipitation of proteins with polyethylene glycol. In: Methods of Plasma Protein Fractionation. Academic Press; 1980. pp 57-74
- 9 Casillas G, Simonetti C. Polyvinylpyrrolidone (PVP) - a new precipitating agent for human and bovine factor VIII and fibrinogen. Br J Haematol 1982; 50: 665-672
- 10 Mason EC, Pepper DS, Griffin B. Production of cryoprecipitate of intermediate purity in a closed system thaw-siphon process. Thromb Haemostas 1981; 46: 543-546
- 11 Rotblat F, Tuddenham EG D. Assays based on a haemophilic and an acquired antibody to VIII:C. Thromb Res 1981; 21: 431-445
- 12 Laurell CB. Quantitative estimation of proteins by electrophoresis in agarose gel containing antibodies. Anal Biochem 1966; 15: 45-52
- 13 Gornall AG, Bardawill CJ, David MM. Determination of serum protein by means of the Biuret reaction. J Biol Chem 1949; 177: 751-766
- 14 Owen WG, Wagner RH. Antihemophilic factor. A new method for purification Thromb Res 1972; 1: 71-88
- 15 Zuber T, Morgenthaler JJ. Polyvinylpyrrolidone as a precipitating agent for factor VIII and fibrinogen. Br J Haematol 1982; 52: 517-518
- 16 Alexander B, Odake K, Lawler D, Swanger M. Coagulation, hemostasis and plasma expanders: a quarter century enigma. Fed Proc 1975; 34: 1429-1440
- 17 Alexander B. Effects of plasma expanders on coagulation and haemostasis: dextran, hydroxyethyl starch and other macromolecules revisited. In: Blood Substitutes and Plasma Expanders. A. R. Liss; New York: 1978. pp 293-326
- 18 Laurent TC. The interaction between polysaccharides and other macromolecules. The solubility of proteins in the presence of dextran Biochem J 1963; 89: 253-257
- 19 Juckes IR M. Fractionation of proteins and viruses with polyethylene glycol. Biochim Biophys Acta 1971; 229: 535-546