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Thromb Haemost 1983; 50(04): 860-863
DOI: 10.1055/s-0038-1665329
Original Article
Schattauer GmbH Stuttgart

A Rapid One-Step Immunoradiometric Assay for Factor VIII-Procoagulant Antigen Utilizing Monoclonal Antibodies

H V Stel
The Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Amsterdam, The Netherlands
,
E C I Veerman
The Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Amsterdam, The Netherlands
,
J G Huisman
The Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Amsterdam, The Netherlands
,
M C Janssen
The Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Amsterdam, The Netherlands
,
J A van Mourik
The Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Amsterdam, The Netherlands
› Author Affiliations
Further Information

Publication History

Received 13 September 1983

Accepted 05 October 1983

Publication Date:
18 July 2018 (online)

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Summary

A two-site immunoradiometric assay for factor VIII-procoagulant antigen (VIIICAg) that relies completely on monoclonal antibodies has been developed. By selecting an appropriate combination of these antibodies, it was possible to develop an assay in which the radiolabelled monoclonal antibody did not inhibit the binding of antigen to the solid-phase monoclonal antibodies. Thus, the entire test could be carried out as a one-step procedure. With this one-step assay, an amount of 0.0025 U VIIICAg/ml plasma could be detected after 4 hr of incubation, whereas 18 hr of incubation resulted in a lower limit of sensitivity of 0.0005 U VIIICAg/ml. The use of a one-step assay provides a significant advantage over the conventional two-step assay by simplifying, shortening and rendering the performance of the assay more convenient.

Keywords

Monoclonal antibodies - One-step IRMA - VIIICAg - Haemophilia A
 

  • References

  • 1 Hoyer LW. The factor VIII complex: structure and function. Blood 1981; 58: 1-13
    PubMedGoogle Scholar
  • 2 Lazarchick J, Hoyer LW. Immunoradiometric measurement of the factor VIII procoagulant antigen. J. Clin Invest 1978; 62: 1048-1052
    CrossrefPubMedGoogle Scholar
  • 3 Peake IR, Bloom AL, Giddings JC, Ludlam CA. An immunoradiometric assay for procoagulant factor VIII antigen: results in haemophilia, von Willebrand’s disease and fetal plasma and serum. Br J Haematol 1979; 42: 269-281
    CrossrefPubMedGoogle Scholar
  • 4 Holmberg L, Borge L, Nilsson R, Nilsson IM. Measurement of antihaemophilic factor antigen (VIII :CAg) with a solid phase immunoradiometric method based on homologous non-haemophilic antibodies. Scand J Haematol 1979; 23: 17-24
    PubMedGoogle Scholar
  • 5 Hellings JA, van LeeuwenF, Over J, van MourikJ. Immunoradiometric assay of VIII :CAg, a potential tool to detect human VIII: C antibodies. Thromb Res 1982; 26: 297-302
    CrossrefPubMedGoogle Scholar
  • 6 Thomas KB, Howard MA, Koutts J, Firkin BG. Simplified immunoradiometric assay for factor VIII coagulant antigen. Br J Haematol 1982; 51: 47-57
    CrossrefPubMedGoogle Scholar
  • 7 Muller HP, van Tilburg N, Derks J, Klein E, Bertina RM. A monoclonal antibody to VIII: C produced by a mouse hybridoma. Blood 1981; 58: 1000-1006
    PubMedGoogle Scholar
  • 8 Brown J, Le Phuc Thuy. 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
  • 9 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
  • 10 Köhler G, Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 1975; 256: 495-497
    CrossrefPubMedGoogle Scholar
  • 11 Wands JR, Carlson RF, Schoemaker H, Isselbacker KJ, Zurawski VR. Immunodiagnosis of hepatitis B with high-affinity IgM monoclonal antibodies. Proc Natl Acad Sei USA 1981; 78: 1214-1218
    CrossrefPubMedGoogle Scholar
  • 12 Goodall AH, Meek FL, Water JA, Miescher GC, Janossy G, Thomas HC. A rapid one-step radiometric assay for hepatitis B surface antigen utilizing monoclonal antibodies. J Immunol Methods 1982; 52: 167-174
    CrossrefPubMedGoogle Scholar
  • 13 Hunter WM, Bennie JG, Brock DJ H, van Heijningen V. Monoclonal antibodies for use in an immunoradiometric assay for α-foetoprotein. J Immunol Methods 1982; 50: 133-144
    CrossrefPubMedGoogle Scholar
  • 14 Stel HV, van der Kwast ThH, Veerman EC I. Detection of Factor VIII coagulant antigen in human liver tissue. Nature 1983; 303: 530-532
    CrossrefPubMedGoogle Scholar
  • 15 van Mourik J, Mochtar IA. Purification of human antihemophilic factor (factor VIII) by gelchromatography. Biochim Biophys Acta 1970; 221: 677-679
    CrossrefPubMedGoogle Scholar
  • 16 Tuddenham EG D, Trabold NC, Collins JA, Hoyer LW. The properties of factor VIII coagulant activity prepared by immunoadsorbent chromatography. J Lab Clin Med 1979; 93: 40-53
    PubMedGoogle Scholar
  • 17 Goding JW. Conjugation of antibodies with fluorchromes: modifica-tion to the standard methods. J Immunol Methods 1976; 13: 215-226
    CrossrefPubMedGoogle Scholar
  • 18 Devare SG, Stephenson JR. Biochemical and immunological characterization of major envelope glycoprotein of bovine leukemia virus. J Virol 1977; 23: 443-447
    PubMedGoogle Scholar
  • 19 Veitkamp JJ, Drion EF, Loeliger EA. Detection of the carrier state in hereditary coagulation disorders I. Thrombos Diathes Haemorrh 1968; 19: 279-303
    PubMedGoogle Scholar
  • 20 Girma JP, Lavergne JM, Meyer D, Larrieu MJ. Immunoradiometric assay of factor VIII: coagulant antigen using four human antibodies. Study of 27 cases of haemophilia A Br J Haematol 1981; 47: 269-282
    CrossrefPubMedGoogle Scholar