Affinity Purification of Plasma Proteins: Characterization of Six Affinity Matrices and their Application for the Isolation of Human Factor VIII

Summary

For the purification of coagulation factor VIII, (1,1’-carbonyldiimida zole [CDI] -activated) Sepharose CL-48 was functionalized with two aminoalkyl and four aminoalkyl-carbamylalkyl ligand- spacer combinations. The affinity matrices were contacted with human plasma. All affinity matrices showed complete adsorption of factor VIII (>90%) and three aminoalkyl-carbamylalkyl Sepharoses gave factor-VIII recoveries of 50-65% and a factor-VIII preparation with a specific activity of 1–2 U factor VIII/mg of protein. Furthermore, no fibrinogen, immunoglobulin G and albumin could be detected in the isolated factor VIII. Optimal results were obtained using the di-methyl-aminopropyl-carbamylpentyl-Sepharose affinity matrix.

Keywords

Factor VIII - Affinity chromatography

• References

• 1 Pool JG, Hershgold EJ, Pappenhagen AR. High potency anti- haemophilic factor concentrate prepared from cryoglobulin precipitate. Nature 1964; 203: 312
• 2 Pool JG, Shannon AE. Production of high potency concentrates of anti-haemophilic globulin in a closed bag system. New Eng J Med 1965; 273: 1443-1447
• 3 Margolis J, Gallovich CM, Rhoades P. Approaches for preparation of “high-purity” factor VIII by controlled pore glass treatment. Vox Sang 1984; 46: 341-348
• 4 Brinkhous KM, Shanbrom E, Roberts HR, Webster WP, Fekete L, Wagner RH. A new high-potency glycine-precipitated antihemophilic factor (AHF) concentrate. J Am Med Assoc 1968; 205: 613-617
• 5 Newman J, Johnson AJ, Karpatkin S, Puszkin S. Methods for the production of clinically effective intermediate- and high-purity factor- VIII concentrates. Br J Haematol 1971; 2: 1-20
• 6 Roberts HR, Macik BG. Factor VIII and IX concentrates: Clinical efficacy as related to purity. In: Thrombosis and Haemostasis 1987. Verstraete M, Vermylen J, Lijnen R, Amout J. eds Leuven University Press; Leuven: 1987. pp 563-581
• 7 Lederman MN, Saunders C, Toosi Z, Lemon N, Everson B, Ratnoff OD. Antihemophiliac factor (FVIII) preparations inhibit lymphocyte proliferation and production of interleukin 2. J Lab Clin Med 1986; 107: 471-478
• 8 Fulcher CA, Zimmerman TS. Characterization of the human factor VIII procoagulant protein with a heterologous precipitating antibody. Proc Natl Acad Sci USA 1982; 79: 1648-1652
• 9 Kuo G, Masiarz FR, Truett M, Valenzuela P, Rasmussen ME, Favaloro J. Monoclonal antibodies to factor VIIIC. US Patent 4: 716-117 1987;
• 10 Hornsey VS, Griffin BD, Pepper DS, Micklem LR, Prowse CV. Immunoaffinity purification of factor VIII complex. Thromb Haemostas 1987; 57: 102-105
• 11 Griffith M, Liu S, Neslund G, Tsang I, Lettelier D, Berkebile R. Preparation of high specific activity plasma AHF by anti-FVIIIc immunoaffinity chromatography. Thromb Haemostas 1987; 58: 307 Abstract
• 12 Miescher PA, Jaffé ER. eds Human factor VIII:c purified using monoclonal antibody to von Willebrand factor. In: Seminars in Hematology 1988; 25 supplement (Suppl. 01) 1-45
• 13 Vukovich Th, Doleschel W, Roller E, Auerswald W. Purification of factor VIII by chromatography on butyl-Sepharose. Proceedings Serono Symposium, 1977 Haemostas Thromb 1979; 15: 407-410
• 14 Austen DE. The chromatographic separation of factor VIII on aminohexyl Sepharose. Br J Haematol 1979; 43: 669-674
• 15 Morgenthaler JJ. Chromatography of antihemophilic factor on diaminoalkane- and aminoalkane-derivatized Sepharose. Thromb Haemostas 1982; 47: 124-127
• 16 Riethorst W, te Booy MP W M, Klumperman L, Ettema JG, Beugeling T, Bantjes A, van Aken WG. Affinity purification of plasma proteins. I. Optimization of the ligand-spacer combination for the isolation of coagulation factor VIII (FVIII) from human blood plasma. Biochemistry. 1988 submitted for publication
• 17 Peterson EA, Sober HA. Chromatography of proteins. I. Cellulose ion-exchange adsorbents. J Am Chem Soc 1956; 78: 751-752
• 18 Veltkamp JJ, Drion EF, Loeliger EA. Detection of the carrier state in hereditary coagulation disorders I. Thromb Diathes Haemorrh 1968; 19: 279-303
• 19 Denson KW E. The specific assay of Prower-Stuart factor and factor VII. Acta Haematol 1961; 15: 105-120
• 20 Biggs R. Human Blood Coagulation. Haemostasis and Thrombosis. 2nd. edition Blackwell Scientific Publications; Oxford: 1976
• 21 Mancini G, Carbonara AO, Heremans JF. Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry 1965; 2: 235-254
• 22 Middleton SM, Bennet IH, Smith JK. A therapeutic concentrate of coagulation factors II, IX and X from citrated factor VUI-deficient plasma. Vox Sang 1973; 24: 441-456
• 23 Heystek J, Brummelhuis HG J, Krijnen HW. Contributions to the optimal use of human blood. II. The large-scale preparation of prothrombin complex. Vox Sang 1973; 25: 113-123
• 24 Faure A, Caron M, Tepenier D. Improved buffer for the chromatographic separation of factor VIII coagulant. J Chromatogr 1983; 257: 387-391
• 25 Neal GG, Hersee D, Smith JK. Purification of factor VIII on aminohexyl Sepharose. Thromb Haemostas 1985; 54: 78 (Abstr)
• 26 Amphlett GW, Hrinda ME. Process for purifying factor VIII:C. US Patent 4, 508, 709, 1985