Download PDF
Thromb Haemost 1988; 59(03): 364-371
DOI: 10.1055/s-0038-1647497
Original Article
Schattauer GmbH Stuttgart

Immunopurification of Human Factor VIII/vWF Complex from Plasma

Odile Mejan
Immunotech, Marseille, France
,
Vincent Fert
Immunotech, Marseille, France
,
Maryléne Delezay
Immunotech, Marseille, France
,
Michel Delaage
Immunotech, Marseille, France
,
Rose Cheballah
Immunotech, Marseille, France
,
Alain Bourgois
Immunotech, Marseille, France
› Author Affiliations
Further Information

Publication History

Received 09 September 1987

Accepted after revision 08 January 1988

Publication Date:
29 June 2018 (online)

Also available at
  PDF Download Permissions and Reprints

Summary

In this study we describe a process for immunopurification of FVIII/vWF complex directly from plasma. A mAb against vWF has been selected that is able to bind, under physiologic conditions, the FVIII/vWF complex and to release it in slightly alkaline conditions while preserving its activity.

After investigating the influence of solid supports and of coupling methods on the recovery of active FVIII we produced an immunoadsorbent by immobilisation of the selected mAb onto a Sephacryl S-1000 support using a benzoquinone coupling method. With this immunoadsorbent we developed a purification process directly from plasma with an excellent recovery (50%) of both FVIII and vWF activities. The product obtained is very enriched (the FVIII: C specific activity is 20 IU/mg of protein) and is stable after lyophilization.

Keywords

Factor VIII - von Willebrand factor - Monoclonal antibodies - Immunopurification - Immunoadsorbent
 

  • References

  • 1 Chavin SI. Factor VIII: structure and function in blood clotting. Am J Hematol 1984; 16: 297-306
    CrossrefPubMedGoogle Scholar
  • 2 Owen WG, Wagner RH. Antihemophilic factor: separation of an active fragment following dissociation by salts or detergents. Thromb Diath Haemorrh 1972; 27: 502-515
    PubMedGoogle Scholar
  • 3 Hoyer LW. Molecular pathology and immunology of factor VIII (hemophilia A and factor VIII inhibitors). Hum Pathol 1987; 18: 153-161
    CrossrefPubMedGoogle Scholar
  • 4 Weiss HJ, Sussman II, Hoyer LW. Stabilization of Factor VIII in plasma by the von Willebrand factor. J Clin Invest 1977; 60: 390-404
    CrossrefPubMedGoogle Scholar
  • 5 Fay PJ, Cavallaro C, Marder VJ. Brief communication: comparison of the in vivo survival of human factor VIII with and without von Willebrand factor in the hemophilic dog. Thromb Res 1986; 37: 425-429
    PubMedGoogle Scholar
  • 6 Gitshier J, Wood WI, Goralka TM, Wion KL, Chen EY, Eaton DH, Vehar GA, Capon DJ, Lawn RM. Characterization of the human factor VIII gene. Nature 1984; 312: 326-330
    CrossrefPubMedGoogle Scholar
  • 7 Toole JJ, Knopf JL, Wozney JM, Sultzman LA, Buecker JL, Pittman DD, Kaufman RJ, Brown E, Shoemaker C, Orr EC, Amphlett WG, Foster WB, Coe ML, Knutson GJ, Fass DN, Hewick RM. Molecular cloning of a cDNA encoding human antihaemophilic factor. Nature 1984; 312: 342-347
    CrossrefPubMedGoogle Scholar
  • 8 Vehar GA, Keyt B, Eaton D, Rodriguez H, O’Brien DP, Rotblat F, Opperman H, Keck R, Wood WI, Hardins RN, Tuddenham EGD, Lawn RM, Capon DJ. Structure of human factor VIII. Nature 1984; 312: 337-342
    CrossrefPubMedGoogle Scholar
  • 9 Wood WI, Capon DJ, Simonsen CC, Eaton DL, Gitshier J, Keyt B, Seeburg PH, Smith DH, Hollingshead P, Wion KL, Delwart E, Tuddenham EGD, Vehar GA, Lawn RM. Expression of active human factor VIII from recombinant DNA clones. Nature 1984; 312: 330-337
    CrossrefPubMedGoogle Scholar
  • 10 Atichartakarn V, Marder VJ, Kirby EP, Budzynski AZ. Effects of enzymatic degradation on the subunit composition and biologic properties of human factor VIII. Blood 1978; 51: 281-297
    PubMedGoogle Scholar
  • 11 Andersson LO, Forsman N, Huang K, Larsen K, Lundin A, Pavlu B, Sandberg H, Sewerin K, Smart J. Isolation and characterization of human Factor VIII: Molecular forms in commercial factor VIII concentrate, cryoprecipitate, and plasma. Proc Natl Acad Sci USA 1986; 83: 2979-2985
    CrossrefPubMedGoogle Scholar
  • 12 Eaton DL, Wood WI, Eaton D, Hass PE, Hollingshead P, Wion K, Mather J, Lawn RM, Vehar GA, Gorman C. Construction and characterization of an active factor VIII variant lacking the central one third of the molecule. Biochemistry 1986; 26: 8342-8347
    PubMedGoogle Scholar
  • 13 Fay PJ, Anderson MT, Chavin SI, Marder VJ. The size of human factor VIII heterodimers and the effects produced by thrombin. Biochim Biophys Acta 1986; 871: 268-278
    CrossrefPubMedGoogle Scholar
  • 14 Sadler JE, Shelton-Inloes BB, Sorace JM, Harlan JM, Titani K, Davie EW. Cloning and characterization of two cDNAs coding for human von Willebrand factor. Proc Natl Acad Sci USA 1985; 82: 6394-6398
    CrossrefPubMedGoogle Scholar
  • 15 Fretto LJ, Fowler WE, Mc Caslin DR, Ericksson HP, Mc Kee PA. Substructure of human von Willebrand Factor. J Biol Chem 1986; 261: 15679-15689
    PubMedGoogle Scholar
  • 16 Furlan M. Factor VIII/von Willebrand factor: a multivalent ligand binding to platelets and collagen. Blut 1986; 52: 329-336
    CrossrefPubMedGoogle Scholar
  • 17 Davies BL, Furlong RA, Peake IR. Studies on the relationship between factor VIII-related antigen (VIII :RAg) and factor VIII clotting antigen (VIII :CAg) by Immunoelectrophoresis and autoradiography using 125I anti-VIII:CAg. Thromb Res 1981; 22: 87-93
    CrossrefPubMedGoogle Scholar
  • 18 Foster PA, Fulcher CA, Marti T, Titani K, Zimmerman TS. A major factor VIII binding domain resides within the amino-terminal 272 amino acid residues of von Willebrand factor. J Biol Chem 1987; 262: 8443-8446
    PubMedGoogle Scholar
  • 19 Pool JG, Shannon AE. Production of high purity concentrates of anti-hemophilic globulin in a closed bag system. N Engl J Med 1965; 273: 1443
    CrossrefPubMedGoogle Scholar
  • 20 Johnson AJ, Karpatkin MH, Newman J. Preparation of and clinical experience with antihemophilic factor concentrates. Thromb Diath Haemorr 1969; suppl (Suppl. 35) 49
    PubMedGoogle Scholar
  • 21 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
    CrossrefPubMedGoogle Scholar
  • 22 Perret BA, Furlan M, Beck EA. Studies on factor VIII-related protein II. Estimation of molecular size differences between factor VIII oligomers. Biochim Biophys Acta 1979; 578: 164
    CrossrefPubMedGoogle Scholar
  • 23 Austen DE G. The chromatographic separation of factor VIII on aminohexyl Sepharose. Br J Haematol 1979; 43: 669-674
    CrossrefPubMedGoogle Scholar
  • 24 Griffith M, Liu S, Neslund G, Tsang I, Lettelier D, Berkebile R. Preparation of high specific activity plasma AHF by anti-FVIII:C immunoaffinitÿ chromatography XIth International Congress on Thrombosis and Haemostasis, Bruxelles. 04.07.1987
    PubMedGoogle Scholar
  • 25 Tuddenham EGD, Trabold NC, Collins JA, Hoyer LW. The properties of factor VIII coagulant activity prepared by immunosorbent chromatography. J Lab Clin Med 1979; 93: 40-53
    PubMedGoogle Scholar
  • 26 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
    CrossrefPubMedGoogle Scholar
  • 27 Stel HV, Veerman ECI, Huisman JG, Janssen MC, Van Mourik JA. A rapid one-step immunoradiometric assay for factor VIII-procoagulant antigen utilizing monoclonal antibodies. Thromb Haemostas 1983; 50: 860-863
    PubMedGoogle Scholar
  • 28 Sultan Y, Avner P, Maisonneuve P, Arnaud D, Jeanneau C. An immunoradiometric assay for factor VIII related antigen (VIIIRAg) using two monoclonal antibodies. Comparison with polyclonal rabbit antibodies for use in von Willebrand’s disease diagnosis Thromb Haemostas 1984; 52: 250-252
    PubMedGoogle Scholar
  • 29 Soulier JP, Larrieu MJ. Nouvelle méthode de diagnostic de l’hémophilie. Dosage des facteurs antihémophiliques A et B. Le sang 1953; 24: 205
    PubMedGoogle Scholar
  • 30 Kirwood TB L, Barrowcliffe TW. Discrepancy between one-stage and two-stage assay of factor VIII :C. Br J Haematol 1978; 40: 333
    CrossrefPubMedGoogle Scholar
  • 31 Howard MA, Firkin BG. Ristocetin, a new tool in the investigation of platelet aggregation. Thromb Diath Haemorrh 1971; 26: 362-369
    PubMedGoogle Scholar
  • 32 Mazurier C, Parquet-Gernez A, Samor B, Goudemand M, Montreuil J. Etude de l’antigène lié au facteur VIII (FVIIIR: Ag) Purification par Chromatographie d’immuno-affinité. C R Acad Sci Paris 1979; 296: 257
    PubMedGoogle Scholar
  • 33 Köhler G, Milstein C. Continuous cultured of fused cell secreting antibody of predefined specificity. Nature 1975; 256: 495
    CrossrefPubMedGoogle Scholar
  • 34 Sola B, Avner P, Sultan Y, Jeanneau C, Maisonneuve P. Monoclonal antibodies against human factor FVIII molecule neutralize antihemophilic factor and ristocetin cofactor activities. Proc Natl Acad Sci 1982; 79: 183-187
    CrossrefPubMedGoogle Scholar
  • 35 Scatchard G. The attraction of proteins to small molecules and ions. Ann N Y Acad Sci 1949; 51: 660
    CrossrefPubMedGoogle Scholar
  • 36 Ey PL, Prowse SJ, Jenkin CR. Isolation of pure IgG1 IgG2a and IgG2b immunoglobulins from mouse serum using protein A-Sepha-rose. Immunochemistry 1978; 15: 429-436
    CrossrefPubMedGoogle Scholar
  • 37 Weston PD, Avrameas S. Proteins coupled to polyacrylamide beads using glutaraldehyde. Biochem Biophys Res Commun 1971; 45: 1574
    CrossrefPubMedGoogle Scholar
  • 38 Axen R, Porath J, Ernback S. Chemical coupling of peptides and proteins to polysaccharides by means of cyanogen halides. Nature 1967; 214: 1302
    CrossrefPubMedGoogle Scholar
  • 39 Cuatrecasas P. Protein purification by affinity chromatography. J Biol Chem 1970; 245: 574
    PubMedGoogle Scholar
  • 40 Porath J, Fornstedt N. Group fractionation of plasma proteins on dipolar ion exchangers. J Chromatogr 1970; 51: 479-489
    CrossrefPubMedGoogle Scholar
  • 41 Nillson K, Mosbach K. Immobilization of enzymes and affinity ligands to various hydroxyl group carrying supports using highly reactive sulfonyl chlorides. Biochem Biophys Res Commun 1981; 102: 449
    CrossrefPubMedGoogle Scholar
  • 42 Finlay TH, Troll V, Levy M, Johnson A, Hodgins LT. New methods for the preparation of biospecific adsorbents and immobilized enzymes utilizing trichloro-s-triazine. Anal Biochem 1978; 87: 77
    CrossrefPubMedGoogle Scholar
  • 43 Brandt J, Andersson LO, Porath J. Covalent attachement of proteins to polysaccharide carriers by means of benzoquinone. Biochim Biophys Acta 1975; 386: 196-202
    CrossrefPubMedGoogle Scholar
  • 44 Brocway W, Fass DN. The nature of the interaction between ristocetin-Willebrand factor and the factor VIII coagulant activity molecule. J Lab Clin Med 1977; 89: 1295-1305
    PubMedGoogle Scholar
  • 45 Hoyer LW. The factor VIII complex: structure and function. Blood 1981; 58: 01-13
    PubMedGoogle Scholar
  • 46 Hornsey VS, Griffin BD, Pepper DS, Micklem LR, Prowse CV. Immunoaffinity purification of factor VIII complex. Thromb Haemostas 1987; 57: 102-105
    PubMedGoogle Scholar
  • 47 Jorieux S, Mazurier C, Deromeuf C, Goudemand M. Properties and interests of a monoclonal antibody to human von Willebrand factor, abstract 14. Bari International Conference on Factor VIII/von Willebrand Factor. 10..1985
    PubMedGoogle Scholar
  • 48 Fass DN, Knutson GJ, Katzmann JA. Monoclonal antibodies to porcine factor VIII coagulant and their use in the isolation of active coagulant protein. Blood 1982; 59: 594-600
    PubMedGoogle Scholar