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Thromb Haemost 1998; 80(04): 624-631
DOI: 10.1055/s-0037-1615433
Rapid Communication
Schattauer GmbH

Modification of Factor VIII in Therapeutic Concentrates after Virus Inactivation by Solvent-Detergent and Pasteurisation

S. Raut
1   From the Division of Haematology, National Institute for Biological Standards and Control, South Mimms, Potters Bar, Hertfordshire, UK; Research and Development Unit, Central Department for Fractionation, Belgian Red Cross, Brussels, Belgium
,
Di M. Giambattista
1   From the Division of Haematology, National Institute for Biological Standards and Control, South Mimms, Potters Bar, Hertfordshire, UK; Research and Development Unit, Central Department for Fractionation, Belgian Red Cross, Brussels, Belgium
,
S. A. Bevan
1   From the Division of Haematology, National Institute for Biological Standards and Control, South Mimms, Potters Bar, Hertfordshire, UK; Research and Development Unit, Central Department for Fractionation, Belgian Red Cross, Brussels, Belgium
,
A. R. Hubbard
1   From the Division of Haematology, National Institute for Biological Standards and Control, South Mimms, Potters Bar, Hertfordshire, UK; Research and Development Unit, Central Department for Fractionation, Belgian Red Cross, Brussels, Belgium
,
T. W. Barrowcliffe
1   From the Division of Haematology, National Institute for Biological Standards and Control, South Mimms, Potters Bar, Hertfordshire, UK; Research and Development Unit, Central Department for Fractionation, Belgian Red Cross, Brussels, Belgium
,
R. Laub
1   From the Division of Haematology, National Institute for Biological Standards and Control, South Mimms, Potters Bar, Hertfordshire, UK; Research and Development Unit, Central Department for Fractionation, Belgian Red Cross, Brussels, Belgium
› Author Affiliations
Further Information

Publication History

Received 01 December 1997

Accepted after resubmission 22 June 1998

Publication Date:
08 December 2017 (online)

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Summary

The addition of a pasteurisation step to a solvent/detergent (SD) treated FVIII concentrate has recently resulted in enhanced inhibitor incidence in patients in Germany and Belgium. We have investigated the effect of virus inactivation procedures on FVIII function by preparing experimental concentrates from the same starting cryoprecipitate with the following procedures: none (N); dry heat (DH); pasteurisation (P); solvent/detergent (SD); solvent detergent + dry heat (SDDH); solvent detergent + pasteurisation (SDP). In addition, several clinical SD concentrates with and without pasteurisation were studied.

There were no significant differences in fibrinogen and vWF content and in the ratio of one-stage/chromogenic FVIII activity among any of the samples studied. In thrombin proteolysis and FXa generation experiments, there were no differences in results on samples N, DH, P, and SDDH from those on sample SD. However sample SDP gave markedly different results from sample SD in the following respects: slower thrombin proteolysis (t½2 = 12.0 min vs 1.9 min); more rapid FXa generation (rate 2.5 times that of SD); enhanced phospholipid binding (KD = 3.89 × 10–11M vs 5.53 × 10–10M). Similar differences between SDP and SD were seen in the clinical samples.

The observed changes in the FVIII activity occurred in combination with SD and pasteurisation, but not with either treatment alone. These results suggest that SDP treatment may enhance exposure of the phospholipid binding site in the C2 domain of FVIII, and since inhibitors to the SDP product are predominantly against C2, these findings could be relevant to the enhanced immunogenicity of the SDP product.

 

  • References

  • 1 Krenz W, Escuriola-Ettingshausen C, Martinez-Saguer Güngör T, Korn-huber B. Epidemiology of inhibitors in haemophilia A. Vox Sang 1996; 70 (suppl): 2-8.
    CrossrefPubMedGoogle Scholar
  • 2 Peerlinck K, Rosendaal FR, Vermylen J. Incidence of inhibitor development in a group of young hemophilia A patients treated exclusively with lyophilized cryoprecipitate. Blood 1993; 81: 3332-5.
    PubMedGoogle Scholar
  • 3 Peerlinck K, Arnout J, Giambattista MD, Gilles JG, Laub R, Jacquemin M, Saint-Remy JMR, Vermylen J. Factor VIII inhibitors in previously treated haemophilia A patients with a double virus-inactivated plasma derived factor VIII concentrate. Thromb Haemost 1997; 77: 80-6.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 4 Rosendaal FR, Nieuwenhuis HK, van den Berg HM, Heijboer H, MauerBunschoten EP, van der Meer J, Smit C, Strengers PFW, Briët E. The Dutch Haemophilia Study Group. A sudden increase in factor VIII inhibitor development in multitransfused haemophilia A patients in the Netherlands. Blood 1993; 81: 2180-6.
    PubMedGoogle Scholar
  • 5 Peerlinck K, Arnout J, Gilles JG, Saint-Remy JMR, Vermylen J. A higher than expected incidence of factor VIII inhibitors in multitransfused haemophilia A patients tested with an intermediate purity pasteurised factor VIII concentrate. Thromb Haemost 1993; 69: 115-8.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 6 Saldanha J, Minor P. Detection of human parvovirus B19 DNA in plasma pools and blood products derived from these pools: implications for efficiency and consistency of removal of B19 DNA during manufacture. Br J Haematol 1996; 93: 714-9.
    CrossrefPubMedGoogle Scholar
  • 7 Vermylen J, Peerlinck K. Review of the hepatitis A epidemics in haemophiliacs in Europe. Vox Sang 1994; 67: 8-11.
    CrossrefPubMedGoogle Scholar
  • 8 Ruymann FB, Krill CE, Halpin TJ, Churchill WH, Ewenstein B, DeMaria A, MancoJohnson MJ, Hoffman RE. Hepatitis A among persons with haemophilia who received clotting factor concentrate. MMWR 1996; 45: 29-32.
    PubMedGoogle Scholar
  • 9 Schwinn H, Stadler M, Josic Dj, Bal F, Gehringer W, Nur I, Schütz R. A solvent/detergent treated, pasteurised and highly purified factor VIII concentrate. Arzneim Forsch/Drug Res 1994; 44: 188-91.
    PubMedGoogle Scholar
  • 10 Laub R, Giambattista MD, Fondu P, Brackmann HH, Lenk H, Scandella D. Restricted epitope specificity for factor VIII inhibitors which appeared in previously treated haemophiliacs after infusion with OCTAVI SD plus. Thromb Haemost 1997; suppl: 590.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 11 Barrowcliffe TW. Inhibitor development and activated factor VIII in concentrates. Thromb Haemost 1993; 70: 1065-6.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 12 Schwinn H, Smith A, Wolter D. Progress in purification of virus-inactivated factor VIII concentrates. Three generations of solvent/detergent treated plasma derivatives. Arzneim Forsch/Drug Res 1989; 39: 1302-5.
    PubMedGoogle Scholar
  • 13 Barrowcliffe TW, Stocks J, Gray E. Preparation of a stable phospholipid reagent for coagulation assays. Haemostasis 1982; 11: 96-101.
    PubMedGoogle Scholar
  • 14 Modi GJ, Blajchman MA, Ofosu FA. The isolation of prothrombin, factor IX and factor X from human FIX concentrates. Thromb Res 1984; 36: 537-47.
    CrossrefPubMedGoogle Scholar
  • 15 Gray E, Walker D, Heath A, Barrowcliffe TW. Collaborative study on assays of activated FIX (FIXa). On behalf of the factor VIII and factor IX subcommittee of the International Society on Thrombosis and Haemostasis (ISTH). Thromb Haemost 1996; 76: 1114-7.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 16 Investigation of a prolonged activated partial thromboplastin time. In: Laboratory Haematology: an account of laboratory technique. Chanarin I. ed. Churchill Livingstone; UK: 1989. pp 293-306.
    Google Scholar
  • 17 Kemball-Cook G, Tubbs JE, Dawson NJ, Barrowcliffe TW. The behaviour of different factor VIII concentrates in a chromogenic factor X-activating system. Brit J Haematol 1993; 84: 273-8.
    CrossrefPubMedGoogle Scholar
  • 18 van Dieijen G, van Rijn JLML, Govers-Riemslag JWP, Hemker HC, Rosing J. Assembly of the intrinsic factor X activating complex: interactions between factor IXa, factor VIIIa and phospholipid. Thromb Haemost 1985; 53: 396-400.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 19 Mertens K, van Wijngaarden A, Bertina RM. The role of factor VIII in the activation of human blood coagulation factor X by activated factor IX. Thromb Haemost 1985; 54: 654-60.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 20 Jönsson U, Fagerstam L, Ivarson B, Johnsson B, Karlsson R, Lundh K, Lofas S, Persson B, Roos H, Ronnberg I, Sjolander S, Stenberg E, Stahlberg R, Urbaniczky C, Ostlin H, Malmquist M. Real-time biospecific interaction analysis using surface plasmon resonance and sensor chip technology. Biotechniques 1991; 11: 620-7.
    PubMedGoogle Scholar
  • 21 Karlsson R, Michaelsson A, Mattsson L. Kinetic analysis of monoclonal antibody-antigen interactions with a new biosensor based analytical system. J Immun Methods 1991; 145: 229-40.
    CrossrefPubMedGoogle Scholar
  • 22 MacDonald RC, MacDonald RI, Menco BPM, Takeshita K, Subbarao NK, Hu LR. Small-volume extrusion apparatus for preparation of large, unilamellar vesicles. Biochim Biophys Acta 1991; 1061: 297-303.
    CrossrefPubMedGoogle Scholar
  • 23 BIAtechnology NOTE 106, BIAcore AB. 1995 pp 1-4.
    PubMedGoogle Scholar
  • 24 BIAcore AB. Evaluating kinetic data. In: BIAevaluation 2.1 Software Handbook. BIAcore AB 1995; pp 4.1-4.24.
    PubMedGoogle Scholar
  • 25 Raut S, Barrowcliffe TW. Interaction of factor VIII with phospholipid (PC:PS 3:1): a kinetic evaluation using surface plasmon resonance (SPR) technology. (In preparation).
    PubMedGoogle Scholar
  • 26 Biggs R, Austen DEG, Denson KWE, Borrett R, Rizza CR. The mode of action of antibodies which destroy factor VIII. II. Antibodies which give complex concentration graphs. Brit J Haematol 1972; 23: 137-55.
    CrossrefPubMedGoogle Scholar
  • 27 Scandella D, Gilbert GE, Shima M, Nakai H, Eagleson C, Felch M, Prescott R, Rajalakshmi KJ, Hoyer LW, Saenko E. Some factor VIII inhibitor antibodies recognize a common epitope corresponding to C2 domain amino acids 2248 through 2312, which overlap a phospholipid-binding site. Blood 1995; 86: 1811-9.
    PubMedGoogle Scholar
  • 28 Prescott R, Nakai H, Saenko EL, Scharrer I, Nilsson IM, Humphries JE, Hurst D, Bray G, Scandella D. The inhibitor antibody response is more complex in hemophilia A patients than in most nonhemophiliacs with factor VIII autoantibodies. Recombinate and Kogenate Study Groups. Blood 1997; 89: 3663-71.
    PubMedGoogle Scholar
  • 29 Hoyer LW, Scandella D. Factor VIII inhibitors: structure and function in autoantibody and hemophilia A patients. Semin Hematol 1994; 31: 1-5.
    PubMedGoogle Scholar
  • 30 Scandella D, Mattingly M, Prescott R. A recombinant factor VIII A2 domain polypeptide quantitatively neutralizes human inhibitor antibodies that bind to A2. Blood 1993; 82: 1767-75.
    PubMedGoogle Scholar
  • 31 Scandella D, Mattingly G, de Graaf S, Fulcher CA. Localisation of epitopes for human factor VIII inhibitor antibodies by immunoblotting and antibody neutralisation. Blood 1989; 74: 1618-26.
    PubMedGoogle Scholar
  • 32 Rosendaal FR. Factor VIII inhibitors on a SD-treated and pasteurized concentrate associated with specific batches and batch characteristics. Thromb Haemost 1997; suppl: 590.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 33 Kemball-Cook G, Edwards SJ, Sewerin K, Andersson LO, Barrowcliffe TW. Factor VIII procoagulant protein interacts with phospholipid vesicles via its 80 kDa light chain. Thromb Haemost 1988; 60: 442-6.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 34 Foster PA, Fulcher CA, Houghten RA, Zimmerman TS. Synthetic factor VIII peptides with amino acids sequences contained within the C2 domain of factor VIII inhibit factor VIII binding to phosphatidylserine. Blood 1990; 75: 1999-2004.
    PubMedGoogle Scholar
  • 35 Josic D, Stadler M, Buchacher A, Schulz P, Schütz R, Wiry G, Schwinn H. Biochemical and immunochemical investigations of different plasma-derived and recombinant factor VIII products. Thromb Haemost 1997; suppl: 512.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 36 Barrowcliffe TW, Dawson NJ, Kemball-Cook G. Activated factor VIII in concentrates prepared by monoclonal antibody and recombinant technology. Thromb Haemost 1989; 62 (1) (suppl): 198.
    PubMedGoogle Scholar
  • 37 Tubbs JE, Kemball-Cook G, Barrowcliffe TW. Effects of von Willebrand factor and phospholipid on co-factor activity of factor VIII in concentrates. Thromb Haemost 1991; 65 (6) (suppl): 944.
    PubMedGoogle Scholar
  • 38 Hill-Eubanks DC, Lollar P. von Willebrand factor is a cofactor for thrombin-catalyzed cleavage of the factor VIII light chain. J Biol Chem 1990; 265 (29) 17854-8.
    PubMedGoogle Scholar
  • 39 Saenko EL, Scandella D. A mechanism for inhibition of factor VIII binding to phospholipid by von Willebrand factor. J Biol Chem 1995; 270: 13826-33.
    CrossrefPubMedGoogle Scholar
  • 40 Yee TT, Williams MD, Hill FGH, Lee CA, Pasi KJ. Absence of inhibitors in previously untreated patients with severe haemophilia A after exposure to a single intermediate purity factor VIII product. Thromb Haemost 1997; 78: 1027-9.
    Article in Thieme ConnectPubMedGoogle Scholar