Using a Monoclonal Antibody to Identify Patients with Type I and Type II von Willebrand’s Disease

Jeffrey D Hall; Dean W Willis; Bruce L Evatt; Debra W Jackson

doi:10.1055/s-0038-1651128

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1987; 57(03): 332-336
DOI: 10.1055/s-0038-1651128

Original Article

Schattauer GmbH Stuttgart

Using a Monoclonal Antibody to Identify Patients with Type I and Type II von Willebrand’s Disease

Jeffrey D Hall

The U. S. Department of Health and Human Services, Public Health Service, Centers for Disease Control, Center for Infectious Diseases, Division of Host Factors, Atlanta, GA, USA

,

Dean W Willis

The U. S. Department of Health and Human Services, Public Health Service, Centers for Disease Control, Center for Infectious Diseases, Division of Host Factors, Atlanta, GA, USA

,

Bruce L Evatt

The U. S. Department of Health and Human Services, Public Health Service, Centers for Disease Control, Center for Infectious Diseases, Division of Host Factors, Atlanta, GA, USA

,

Debra W Jackson

The U. S. Department of Health and Human Services, Public Health Service, Centers for Disease Control, Center for Infectious Diseases, Division of Host Factors, Atlanta, GA, USA

› Author Affiliations

Abstract

PDF Download

Summary

Three monoclonal antibodies produced against vWF:Ag by conventional hybridoma technique did not inhibit factor VIII coagulant activity (F.VIII:C) but did inhibit VIII ristocetin cofactor activity. The antibodies were used in an indirect competitive ELISA for quantifying von Willebrand’s antigen (vWF: Ag) and compared with values obtained by the Laurell technique using commercial antibody by means of a ratio: ELISA/Laurell. For one monoclonal BD2-CC9, vWF:Ag values obtained in the two assays were in good agreement for normal and hemophilia A plasmas (normal, n = 19, ratio = 1.13 ± .17, hemophilia A, n = 10, ratio = 0.91 ± .15). However, type II vWD patients had a disproportionately low value of vWF: Ag with the ELISA. Use of the ratio normalized the difference among individual plasma values and allowed a significant separation of type II vWD plasma (n = 9, ratio = 0.46 ± .19) from normal plasma (p = .0001) and type I vWD plasma (n = 8, ratio = 1.52 ± .34) from type II vWD plasma (p = .0003) using BD2-CC9. Although the sample size was small, the greater degree of discrimination among the vWD plasmas tested with BD2-CC9 (compared with the other two antibodies [CA3-AE4, CC6-BG10]) suggests that this antibody may recognize conformational epitopes that reflect the degree of multimeric polymerization of the vWF molecule rather than simply recognize a decreased number of antigenic sites in a basic subunit. BD2-CC9 may be valuable in investigating the various types of vWD and/or the process of polymerization of this complex protein.

Keywords

Monoclonal antibody - Factor VIII - von Willebrand factor - ELISA - Factor VIII related antigen

PDF (1228 kb)

References

References
1 Ruggeri ZM, Zimmerman TS. Variant von Willebrand’s disease. Characterization of two subtypes by analysis of multimeric composition of factor VIII/von Willebrand factor in plasma and platelets. J Clin Invest 1980; 65: 1318-1325
2 Zimmerman TS, Ruggeri ZM, Fulcher CA. Factor VIII/von Willebrand factor. pp 279-309 In: Brown EB. (ed.) Progress in hematology. volume XIII Grune and Stratton; New York, NY: 1983
3 Ey PL, Prowse SJ, Jenkin CR. Isolation of pure IgG₁ IgG_2a and IgG_2b immunoglobulins from mouse serum using Protein A-Sepharose. Immunochemistry 1978; 15: 429-436
4 Sibley CA. Thrombosis and hemostasis procedures. U. S. Department of Health and Human Services, Centers for Disease Control; Atlanta, GA: 1976
5 Sola B, Avner P, Sultan Y, Jeanneau C, Maisonueuve P. Monoclonal antibodies against human factor VIII molecule neutralize antihemophilic factor and ristocetin cofactor activities. Proc Natl Acad Sci USA 1982; 79: 183-187
6 Bradley LA, Franco EL, Riesner HM. Use of monoclonal antibodies in an enzyme immunoassay for factor VIII-related antigen. Clin Chem 1984; 30: 87-92
7 Ruggeri ZM, Zimmerman TS. The complex multimeric composition of factor VIII/von Willebrand factor. Blood 1981; 57: 1140-1143
8 Tsang V CW, Peralta JM, Simons AR. Enzyme-linked immunoelectrotransfer blot techniques (EITB) for studying the specificities of antigens and antibodies separated by gel electrophoresis. Meth Enzymology 1983; 92: 377-391
9 Snedecor GW, Cochran WG. Statistical methods. The Iowa State University Press; 1980
10 Lehmann EL. Nonparametrics: Statistical methods based on ranks. Holden-Day, Inc.; 1975
11 Stel HV, Sakariassen S, Scholte BJ, Veerman E CI, Vander Kwast TH, De Groot PG, Sixma JJ, Van Mourik JA. Characterization of 25 monoclonal antibodies to factor VUI-von Willebrand factor: relationship between ristocetin-induced platelet aggregation and platelet adherence to subendothelium. Blood 1984; 63: 1408-1415
12 Ardaillou N, Girma JP, Meyer D, Lavergne JM, Shoa’i I, Larrieu MJ. “Variants” of von Willebrand disease. Demonstration of a decreased antigenic reactivity by immunoradiometric assay. Thromb Res 1978; 12: 817-830
13 Girma JP, Ardaillou N, Meyer D, Lavergne JM, Larrieu MJ. Fluidphase immunoradiometric assay for the detection of qualitative abnormalities of factor VIII/von Willebrand factor in variants of von Willebrand’s disease. J Lab Clin Med 1979; 93: 926-939
14 Girma JP, Peitu G, Lavergne JM, Meyer D, Larrieu MJ. Abnormal antigenic reactivity of factor VIII/von Willebrand factor subunit in variants of von Willebrand’s disease. J Lab Clin Med 1982; 99: 481-494
15 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
16 Short PE, Williams CE, Enayat MS, Picken AM, Hill F GH. 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
17 Weis JW, Nelson D. Factor VIII related antigen in variant von Willebrand’s disease. Thromb Res 1984; 33: 457-459
18 Nilsson IM, Peake IR, Bloom AL, Meyer D, Veltkamp J, Green D. The relationship between ristocetin co-factor activity (VIIIR:Cof) and factor related antigen (VIIIR:Ag). Thromb Haemostas 1980; 43: 167-168
19 Thomas JE, Peake IR, Giddings JC, Welch AN, Bloom AL. The application of a monoclonal antibody to factor VIII related antigen (VIIIRAg) in immunoradiometric assays for factor VIII. Thromb Haemostas 1985; 53: 143-147