Download PDF
Thromb Haemost 1986; 55(01): 040-046
DOI: 10.1055/s-0038-1661443
Original Article
Schattauer GmbH Stuttgart

The Production and Characterisation of a Panel of Ten Murine Monoclonal Antibodies to Human Procoagulant Factor VIII

B D Griffin
1   The Scottish National Blood Transfusion Service Headquarters Unit Laboratory, Edinburgh, Scotland, UK
,
L R Micklem
2   The Department of Surgery, University of Edinburgh Medical School, Edinburgh, Scotland, UK
,
M C McCann
2   The Department of Surgery, University of Edinburgh Medical School, Edinburgh, Scotland, UK
,
K James
2   The Department of Surgery, University of Edinburgh Medical School, Edinburgh, Scotland, UK
,
D S Pepper
1   The Scottish National Blood Transfusion Service Headquarters Unit Laboratory, Edinburgh, Scotland, UK
› Author Affiliations
Further Information

Publication History

Received 05 August 1985

Accepted 14 October 1985

Publication Date:
19 July 2018 (online)

  PDF Download  Permissions and Reprints

Summary

A panel of 10 murine monoclonal antibodies to procoagulant FVIII has been developed from the fusion of a single spleen. Balb/c mice were injected with a purified preparation of FVIII: Ag, and antibody production in sera and hybrid culture supernatants was monitored using a specific radiometric screening assay. The antibodies all inhibit FVIII clotting activity in normal plasma, and when immobilised on agarose retain their ability to recognise and bind the FVIII procoagulant protein. Studies on protein A-purified immunoglobulins demonstrate a range of properties within the panel of antibodies with regard to species crossreactivity, clotting inhibition and immunoadsorption. The panel of antibodies has been used to screen heat-treated FVIII concentrates for the occurrence of heat-induced neoantigens.

Key words

Factor VIII - Monoclonal antibodies
 

  • References

  • 1 Hoyer LW. The factor VIII complex: structure and function. Blood 1981; 58: 1-13
    PubMedGoogle Scholar
  • 2 Shapiro SS, Hultin MB. Acquired inhibitors to the blood coagulation factors. Sem Thromb Haemostas 1975; 1-336
    PubMedGoogle Scholar
  • 3 Fay PJ, Chavin SI, Schroeder D, Young FE, Marder VJ. Purification and characterisation of a highly purified human factor VIII consisting of a single type of polypeptide chain. Proc Nat Acad Sci USA 1982; 79: 7200-720
    CrossrefPubMedGoogle Scholar
  • 4 Tran TH, Marbet GA, Duckert F. Rabbit antibodies against the procoagulant activity (VIII :C) of human factor VIII. Thromb Haemostas 1981; 46: 699-705
    PubMedGoogle Scholar
  • 5 Kohler G, Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 1975; 256: 495-497
    CrossrefPubMedGoogle Scholar
  • 6 Sola B, Anver P, Sultan Y, Jeanneau C, Maisonneuve P. Monoclonal antibodies against the human factor VIII molecule neutralise antihemophilic factor and ristocetin cofactor activities. Proc Nat Acad Sci USA 1982; 79: 183-187
    CrossrefPubMedGoogle Scholar
  • 7 Brown JE, Thuy LP, Carton CL, Hougie C. Studies on a monoclonal antibody to human factor VIII coagulant activity, with a description of a facile two-site factor VIII coagulant antigen assay. J Lab Clin Med 1983; 101: 793-805
    PubMedGoogle Scholar
  • 8 Laurell CB. Quantitative estimation of proteins by electrophoresis in agarose gel containing antibodies. Anal Biochem 1966; 15: 45-52
    CrossrefPubMedGoogle Scholar
  • 9 Rotblat F, Tuddenham EG D. Immunologic studies of factor VIII coagulant activity (VIII :C). I. Assays based on a haemophilic and an acquired antibody to VIII :C. Thromb Res 1981; 21: 431-545
    CrossrefPubMedGoogle Scholar
  • 10 Tuddenham EG D, Lane RS, Rotblat F, Johnson AJ, Snape TJ, Middleton S, Kernoff PB A. Response to infusions of poly electrolyte-fractionated human factor VIII concentrate in human haemophilia A and von Willebrand’s disease. Br J Haematol 1982; 52: 259-267
    CrossrefPubMedGoogle Scholar
  • 11 Stel HV, Veerman EC I, Huisman JG, Janssen MC, van Mourik JA. A rapid one-step immunoradiometric assay for factor VIII procoagulant antigen utilising monoclonal antibodies. Thromb Haemostas 1983; 50: 860-86
    PubMedGoogle Scholar
  • 12 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
    PubMedGoogle Scholar
  • 13 Wright JF, Hunter WM. A convenient replacement for cyanogen bromide-activated solid phases in immunoradiometric assays. J Immunol Method 1982; 48: 311-32
    CrossrefPubMedGoogle Scholar
  • 14 Salacinski PR R, McLean C, Sykes JE C, Clement-Jones VV, Lowry PJ. Iodination of proteins, glycoproteins and peptides using a solid phase oxidising agent, 1,3,4,6-tetrachloro 3α, 6α-diphenylglycouril (Iodogen). Anal Biochem 1981; 117: 136-146
    CrossrefPubMedGoogle Scholar
  • 15 Greenwood FC, Hunter WM, Glover JS. The preparation of 131I-labelled human growth hormone of high specific radioactivity. Biochem J 1963; 89: 114-123
    CrossrefPubMedGoogle Scholar
  • 16 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
  • 17 Hunter WM. Simplified solid phase radioimmunoassay without centrifugation. Acta Endocrinol 1977; 85 (212) 463
    PubMedGoogle Scholar
  • 18 Kasper CK, Aldedort LM, Counts RB, Edson JR, Fratantoni J, Green D, Hampton JW, Hilgartner MW, Lazerson J, Levine PH, McMillan CW, Pool JG, Shapiro SS, Sulman NR, Van Eys J. A more uniform measurement of factor VIII inhibitors. Thromb Diath Haemorrh 1975; 34: 869
    PubMedGoogle Scholar
  • 19 Kohler G, Howe SC, Milstein C. Fusion between immunoglobulin-secreting and non secreting myeloma cell lines. Eur J Immunol 1976; 6: 292-295
    CrossrefPubMedGoogle Scholar
  • 20 Kennett RH, Denis KA, Tung AS, Klinman NR. Hybrid plasmacytoma production: fusions with adult spleen cells, monoclonal spleen fragments, neonatal spleen cells and human spleen cells. Curr Topics Microbiol Immunol 1978; 81: 77-91
    PubMedGoogle Scholar
  • 21 Oi V, Herzenberg LA. Immunoglobulin producing hybrid cell lines. In: Selected Methods in Cellular Immunology Mishell BB, Shiigi SM. (eds) W H Freeman & Co; San Francisco: 1980: 351-372
  • 22 Ey PL, Prowse SJ, Jenkins CR. Isolation of pure IgG1, IgG2a and IgG2b immunoglobulins from mouse serum using protein A Sephar-ose. Immunochem 1978; 15: 429-436
    CrossrefPubMedGoogle Scholar
  • 23 Kohn J, Wilchek M. A new approach (cyano-transfer) for cyanogen bromide activation of Sepharose at neutral pH, which yields activated resins, free of interfering nitrogen derivatives. Biochem Biophys Res Commun 1982; 107: 878-884
    CrossrefPubMedGoogle Scholar
  • 24 Tuddenham EG D, Trabold NC, Hoyer LW. The properties of factor VIII coagulant activity prepared by immunoadsorbent chromatography. J Lab Clin Med 1979; 93: 40-53
    PubMedGoogle Scholar
  • 25 Rotblat F, Hawkey C, O’Brien DP, Tuddenham EG D. Immunologic studies of factor VIII coagulant activity (VIII :C). 2. Factor VIII in selected vertebrates. Thromb Res 1982; 25: 423-431
    CrossrefPubMedGoogle Scholar
  • 26 Bier M, Foster PR. Purification of antihemophilia factor VIII by precipitation with zinc ions. U S Patent 4, 406, 886. Sept 27 1983
    PubMedGoogle Scholar
  • 27 Rotblat F, Goodall AH, O’Brien DP, Rawlings E, Middleton S, Tuddenham EG D. Monoclonal antibodies to human procoagulant factor VIII. J Lab Clin Med 1983; 101: 736-746
    PubMedGoogle Scholar
  • 28 Muller HP, van Tilburg NH, Derks J, Klein-Breteler E, Bertina RM. A monoclonal antibody to VIII: C produced by a mouse hybridoma. Blood 1981; 58: 1000-1006
    PubMedGoogle Scholar
  • 29 Gawryl MS, Hoyer LW. Inactivation of FVIII coagulant activity by two different types of human antibodies. Blood 1982; 60: 1103-1109
    PubMedGoogle Scholar
  • 30 Vehar GA, Keyt B, Eaton D, Rodriguez H, O’Brien DP, Rotblat F, Oppermann H, Keck R, Wood WI, Harkins RN, Tuddenham EG D, Lawn RM, Capon DJ. Structure of human factor VIII. Nature 1984; 312: 337-342
    CrossrefPubMedGoogle Scholar
  • 31 Moake JL, Weinstein MJ, Troll JH, Chute LE, Colannino NM. Direct radioimmune detection in human plasma of the association between factor VIII procoagulant protein and von Willebrand factor, and the interaction of von Willebrand factor-bound procoagulant VIII with platelets. Blood 1983; 61: 1167-1173
    PubMedGoogle Scholar
  • 32 Joshua DE, Exner T, Holmes D, Rickard KA, Kronenberg H. Antibodies to human factor VIII complex from Balb/c mice. Thromb Res 1981; 22: 345-352
    CrossrefPubMedGoogle Scholar
  • 33 Bird AG, Codd AA, Collins A. Haemophilia and AIDS. Lancet 1985; 1: 162-163
    PubMedGoogle Scholar