Download PDF
Thromb Haemost 1988; 59(02): 251-254
DOI: 10.1055/s-0038-1642764
Original Articles
Schattauer GmbH Stuttgart

The Application of the Biotin/Avidin System to the von Willebrand Factor Antigen Immunoassay

L D Taylor
The Department of Clinical Haematology and Oncology, Royal Children's Hospital, Melbourne, Australia
› Author Affiliations
Further Information

Publication History

Received 14 September 1987

Accepted after revision 03 December 1987

Publication Date:
21 May 2018 (online)

  PDF Download  Permissions and Reprints

Summary

A vWf: Ag ELISA which uses biotinylated anti-vWf: Ag IgG and an avidin/biotinylated peroxidase (ABP) complex detection system has been developed. The sensitivity of the ELISA for vWf: Ag was 4 × 10-5 U/ml which is approximately 10-fold lower than previously reported vWf: Ag ELISA’s. The avidin used for the preparation of the ABP complex was glutaraldehyde-treated. This treatment increased the reactivity 2-3-fold as compared to ABP complex prepared using untreated avidin. The reactivity of two commercial detection reagents was also tested.

The effect of the reduction of the total assay time to 3.5 h is also described. In this form the detection limit for vWf: Ag was 2 × 10-4 U/ml, the intra-assay precision at vWf: Ag concentrations of 0.01 and 0.0025 U/ml was 3.8% and 4.9% respectively and ELISA results for normal individuals still correlated well with those obtained using electro-immunodiffusion (r = 0.93).

Keywords

Von Willebrand’s disease - ELISA - vWf: Ag - Avidin/biotin system
 

  • References

  • 1 Hoyer LW. The factor VIII complex: structure and function. Blood 1981; 58: 1-13
    PubMedGoogle Scholar
  • 2 Short PE, Williams CE, Picken AM, Hill FG H. Factor VIII related antigen: an improved enzyme immunoassay. Med Lab Sci 1982; 39: 351-355
    PubMedGoogle Scholar
  • 3 Wang HX, George J, Thorpe GH G, Stott RA, Kricka LJ, Whitehead TP. Enhanced luminescence enzyme immunoassay for factor VIII related antigen. J Clin Pathol 1985; 38: 317-319
    CrossrefPubMedGoogle Scholar
  • 4 Silveira AM V, Yamamoto T, Adamson L, Hessel B, Blomback B. Application of an enzyme-linked immunosorbent assay (ELISA) tovon Willebrand factor (vWf) and its derivatives. Thromb Res 1986; 43: 91-102
    CrossrefPubMedGoogle Scholar
  • 5 Ingerslev J. A sensitive ELISA for von Willebrand factor (vWf: Ag). Scand J Clin Lab Invest 1987; 47: 143-149
    CrossrefPubMedGoogle Scholar
  • 6 Zimmerman TS, Hoyer LW, Dickson L, Edgington TS. Determination of the von Willebrand’s disease antigen (factor VUI-related antigen) in plasma by quantitative immunoelectrophoresis. J Lab Clin Med 1975; 86: 152-159
    PubMedGoogle Scholar
  • 7 Ardaillou N, Girma JP, Meyer D, Lavergne JM, Shoa’i I, Larrieu MJ. “Variants” of von Willebrand’s disease. Demonstration of a decreased antigenic reactivity by immunoradiometric assay. Thromb Res 1978; 12: 817-30
    CrossrefPubMedGoogle Scholar
  • 8 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
    CrossrefPubMedGoogle Scholar
  • 9 Girma JP, Ardaillou N, Meyer D, Lavergne JM, Larrieu MJ. Fluid- phase immunoradiometric assay for the detection of qualitative abnormalities of factor VUI/von Willebrand factor in variants of von Willebrand’s disease. J Lab Clin Med 1979; 93: 926-939
    PubMedGoogle Scholar
  • 10 Wilchek M, Bayer EA. The avidin-biotin complex in immunology. Immunol Today 1984; 5: 39-43
    CrossrefPubMedGoogle Scholar
  • 11 Kendall C, Ionescu-Matiu I, Dreesman GR. Utilization of the biotin/ avidin system to amplify the sensitivity of the enzyme-linked immunosorbent assay (ELISA). J Immunol Methods 1983; 56: 329-339
    CrossrefPubMedGoogle Scholar
  • 12 Guesdon J-L, Ternynck T, Avrameas S. The use of avidin-biotin interaction in immunoenzymatic techniques. J Histochem Cytochem 1979; 27: 1131-1139
    CrossrefPubMedGoogle Scholar
  • 13 Hsu S-M, Raine L, Fanger H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J His-tochem Cytochem 1981; 29: 577-580
    CrossrefPubMedGoogle Scholar
  • 14 Hsu S-M, Raine L, Fanger H. A comparative study of the peroxidase- antiperoxidase method and an avidin-biotin complex method for studying polypeptide hormones with radioimmunoassay antibodies. Am J Clin Pathol 1981; 75: 734-738
    CrossrefPubMedGoogle Scholar
  • 15 Dowling SV, Muntz RH, D’souza S, Ekert H. Platelet release abnormality associated with a variant of von Willebrand’s disease. Blood 1976; 47: 265-274
    PubMedGoogle Scholar
  • 16 Zimmerman TS, Abildgaard CF, Meyer D. The factor VIII abnormality in severe von Willebrand’s disease. N Engl J Med 1979; 301: 1307-1310
    CrossrefPubMedGoogle Scholar
  • 17 Italian working group. Spectrum of von Willebrand’s disease: a study of 100 cases. Br J Haematol 1977; 35: 101-112
    CrossrefPubMedGoogle Scholar
  • 18 Girma JP, Pietu G, Lavergne JM, Meyer D, Larrieu MJ. Abnormal antigenic reactivity of factor VUI/von Willebrand factor subunit in variants of von Willebrand’s disease. J Lab Clin Med 1982; 99: 481-494
    PubMedGoogle Scholar
  • 19 Weiss HJ, Pietu G, Rabinowitz R, Girma JP, Rogers J, Meyer D. Heterogeneous abnormalities in the multimeric structure, antigenic properties, and plasma-platelet content of factor VUI/von Willebrand factor in subtypes of classic (type I) and variant (type IIA) von Willebrand’s disease. J Lab Clin Med 1983; 101: 411-425
    PubMedGoogle Scholar
  • 20 Short PE, Williams CE, Enayat MS, Picken AM, Hill FG H. Lack of correlation between factor VIII related antigen, multimeric analysis pattern and parallel or non-parallel dose response curves in an ELISA factor VIII related antigen assay. J Clin Pathol 1984; 37: 194-199
    CrossrefPubMedGoogle Scholar
  • 21 Howard MA, Hau L, Perkin J, Thomas KB, Firkin BG, Koutts J. Causes for the discrepancies in the measurements of factor VIII antigen. Thromb Res 1980; 19: 63-72
    CrossrefPubMedGoogle Scholar
  • 22 Meyer D, Obert B, Pietu G, Lavergne JM, Zimmerman TS. Multimeric structure of factor VUI/von Willebrand factor in von Willebrand’s disease. J Lab Clin Med 1980; 95: 590-602
    PubMedGoogle Scholar
  • 23 Weis JW, Nelson D. Factor VIII related antigen in variant von Willebrand’s disease. Thromb Res 1984; 33: 457-459
    CrossrefPubMedGoogle Scholar
  • 24 Johnson DA, Gautsch JW, Sportsman JR, Elder JH. Improved technique utilising nonfat dry milk for analysis of proteins and nucleic acids transferred to nitrocellulose. Gene Anal Techn 1984; 1: 3-8
    CrossrefPubMedGoogle Scholar
  • 25 Finn FM, Titus G, Montibeller JA, Hofmann K. Hormone receptor studies with avidin and biotinylinsulin-avidin complexes. J Biol Chem 1980; 255: 5742-5746
    PubMedGoogle Scholar
  • 26 Bayer EA, Ben-Hur H, Wilchek M. Enzyme-based detection of glycoproteins on blot transfers using avidin-biotin technology. Anal Biochem 1987; 161: 123-131
    CrossrefPubMedGoogle Scholar