Download PDF
Thromb Haemost 1985; 53(01): 143-147
DOI: 10.1055/s-0038-1661256
Original Article
Schattauer GmbH Stuttgart

The Application of a Monoclonal Antibody to Factor VIII Related Antigen (VIIIRAg) in Immunoradiometric Assays for Factor VIII

J E Thomas
The Department of Haematology, Welsh National School of Medicine, Heath Park, Cardiff, UK
,
I R Peake
The Department of Haematology, Welsh National School of Medicine, Heath Park, Cardiff, UK
,
J C Giddings
The Department of Haematology, Welsh National School of Medicine, Heath Park, Cardiff, UK
,
A N Welch
The Department of Haematology, Welsh National School of Medicine, Heath Park, Cardiff, UK
,
A L Bloom
The Department of Haematology, Welsh National School of Medicine, Heath Park, Cardiff, UK
› Author Affiliations
Further Information

Publication History

Received 16 March 1984

Accepted 02 August 1984

Publication Date:
18 July 2018 (online)

Also available at
  PDF Download  Permissions and Reprints

Summary

A two site solid phase immunoradiometric assay (IRMA) for VIIIRAg has been developed using a monoclonal antibody as both the solid phase ligand and the radiolabelled antibody. A range of normal plasmas and von Willebrand’s disease (vWd) plasmas has been assayed for VIIIRAg by this method and compared with VIIIRAg values measured by an IRMA using polyclonal antibodies and by immunoelectrophoresis and with ristocetin cofactor activity (VIIIRiCoF). It was found that the monoclonal IRMA correlated well with the polyclonal IRMA and the immunoelectrophoretic assay, but the correlation with the VIIIRiCoF assay was relatively poor, in spite of the strong inhibitory effect of the antibody on VIIIRiCoF activity.

Key words

Factor VIII - Monoclonal antibodies - Immunoradiometric assays
 

  • References

  • 1 Bloom AL. The von Willebrand Syndrome. Semin Haematol 1980; 27: 215-227
    PubMedSearch in Google Scholar
  • 2 Hoyer LW. The factor VIII complex: Structure and function. Blood 1981; 58: 1-13
    PubMedSearch in Google Scholar
  • 3 Giddings JC. The purification of factors VIII and IX and the production of specific antisera. In The Haemophilias. Bloom AL. (Ed) Churchill Livingstone; Edinburgh: 1982. pp 551-580
    Search in Google Scholar
  • 4 Kohler G, Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 1975; 256: 495-497
    CrossrefPubMedSearch in Google Scholar
  • 5 Meyer D, Obert B, Zimmermann TS, Edgington T. Monoclonal antibodies specific for factor VIII from cellular hybrids. Circulation 1980; 62 Suppl (Suppl. 111) 106 (Abstr)
    PubMedSearch in Google Scholar
  • 6 Sola B, Avner P, Sultan Y, Jeanneau C, Maisonneuve P. Monoclonal antibodies against the human factor VIII molecule neutralize anti-haemophilic factor and ristocetin cofactor activities. Proc Nat Acad Sci 1982; 79: 183-187
    CrossrefPubMedSearch in Google Scholar
  • 7 Fulcher CA, Zimmerman TS. Characterisation of the human factor VIII procoagulant protein with a heterologous precipitating antibody. Proc Nat Acad Sci 1982; 79: 1648-1652
    CrossrefPubMedSearch in Google Scholar
  • 8 Ogata K, Saito H, Ratnoff OD. The relationship of the properties of antihemophilic factor (Factor VIII) that support Ristocetin-induced platelet agglutination (Factor VIIIR:RC) and platelet retention by glass beads as demonstrated by a monoclonal antibody. Blood 1983; 61: 27-34
    PubMedSearch in Google Scholar
  • 9 Counts RB. A solid-phase immunoradiometric assay of factor VIII protein. Br J Haematol 1975; 31: 429-436
    CrossrefPubMedSearch in Google Scholar
  • 10 Ruggeri ZM, Mannucci PM, Jeffcoate SL, Ingram GI C. Immunoradiometric assay of factor VIII related antigen. With observations in 32 patients with von Willebrand’s disease Br J Haematol 1976; 33: 221-232
    CrossrefPubMedSearch in Google Scholar
  • 11 Peake IR, Bloom AL. The use of an immunoradiometric assay for factor VIII related antigen in the study of atypical von Willebrand’s disease. Thromb Res 1977; 10: 27-32
    CrossrefPubMedSearch in Google Scholar
  • 12 Laurell C. Quantitative estimation of proteins by electrophoresis in agarose gel containing antibodies. Anal Biochem 1966; 15: 45-52
    CrossrefPubMedSearch in Google Scholar
  • 13 Macfarlane DE, Stibbe J, Kirby EP, Zucker MB, Grant RA, McPherson J. A method for assaying von Willebrand factor (Ristocetin cofactor). Thrombos Diathes Haemorrh 1975; 34: 306-308
    PubMedSearch in Google Scholar
  • 14 Greenwood FC, Hunter WM, Glover JS. The preparation of 125I labelled; human growth hormone of high specific radioactivity. Biochem J 1963; 89: 114-123
    CrossrefPubMedSearch in Google Scholar
  • 15 Galfre G, Howe SC, Milstein C, Butcher GW, Howard JC. Antibodies to major histocompatability antigens produced by hybrid cell lines. Nature 1977; 266: 550-552
    CrossrefPubMedSearch in Google Scholar
  • 16 Shearn SA M, Peake IR, Giddings JC, Humphrys J, Bloom AL. The characterisation and synthesis of antigens related to FVIII in vascular endothelium. Thromb Res 1977; 11: 43-56
    CrossrefPubMedSearch in Google Scholar
  • 17 Bloom AL, Giddings JC, Wilks CJ. Factor VIII on the vascular intima possible importance in haemostasis and thrombosis. Nature (New Biology) 1973; 241: 217-219
    CrossrefPubMedSearch in Google Scholar
  • 18 Hudson L, Hay FC. Practical Immunology 2nd Edition. Blackwell Scientific Publications; Oxford: 1983
    Search in Google Scholar
  • 19 Hardisty RM, Mac Pherson JC. A one-stage factor VIII (anti- haemophilic globulin) assay and its use on venous and capillary plasma. Thrombos Diathes Haemorrh 1962; 34: 869-872
    PubMedSearch in Google Scholar
  • 20 Meyer D, Obert B, Pietu G, Lavergne JM, Zimmerman TS. The multimeric structure of factor VIII/von Willebrand factor in von Willebrand’s disease. J Lab Clin Med 1980; 95: 590-602
    PubMedSearch in Google Scholar
  • 21 Furlong BL, Peake IR. An electroblotting technique for the detection of factor VIII/von Willebrand factor multimers in plasma. Br J Haematol 1983; 53: 641-653
    CrossrefPubMedSearch in Google Scholar
  • 22 Bloom AL. Immunological detection of blood coagulation factors in haemorrhagic disorders. Recent Advances in Haematology. Hoff-brand AV, Brain MC, Hirsh J. (Eds) Churchill Livingstone; Edinburgh: 1977. pp 387-411
    Search in Google Scholar
  • 23 Hoyer LW, Rizza CR, Tuddenham EG D, Carta CA, Armitage H, Rotblat F. Von Willebrand factor multimer patterns in von Willebrand’s disease. Br J Haematol 1983; 55: 493-507
    CrossrefPubMedSearch in Google Scholar
  • 24 Counts RB, Paskell SL, Elgee SK. Disulphide bonds and the quaternary structure of factor VIII/von Willebrand factor. J Clin Invest 1978; 62: 702-709
    CrossrefPubMedSearch in Google Scholar