Subscribe to RSS
DOI: 10.1055/s-0038-1660132
Disparity Between One- and Two-Stage Factor VIII Assays in Measuring VIIIC in Heparinized Plasma
Publication History
Received 20 June 1984
Accepted 22 August 1985
Publication Date:
19 July 2018 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/198504/lookinside/thumbnails/10-1055-s-0038-1660132-1.jpg)
Summary
The level of Factor VIII procoagulant activity (VIIIC) was found to be more than 30% higher in heparinized plasma than in citrated plasma from the same donor when determined by a one-stage assay (1.34 U/ml versus 1.0 U/ml). However, after Al(OH)3 adsorption and two-stage assay, the VIIIC levels were not significantly different for the two types of plasma (0.90 ± 0.25 U/ ml versus 0.82 ± 0.23 U/ml). If a one-stage assay was employed after adsorption, a similar low result was obtained (0.92 ± 0.21 U/ ml versus 0.91 ± 0.12 U/ml). This indicated that the adsorption step itself was responsible for the difference in results. Parallel analyses of other VIII markers demonstrated that more VIIIC antigen (VIIICAg) was removed by adsorption of heparinized rather than citrated plasma (0.33 U/ml versus 0.12 U/ml) although there was no difference in the level of VUI-related antigen (VIIIRAg). The results show that adsorption of heparinized plasma prior to either one- or two-stage assay leads to a greater loss of both VIIIC and VIIICAg activity than observed with citrated plasma and indicate that, when applied to heparin plasma the two-stage assay for VIIIC underestimates the actual VIIIC content.
-
References
- 1 Rock GA, Cruickshank WH, Tackaberry ET, Palmer DS. Improved yields of Factor VIII from heparinized plasma. Vox Sang 1979; 36: 294-300
- 2 Rock GA. FVIIIC one-stage assay of plasma containing heparin. In Development systems for efficient separation of Factor VIII from human fresh frozen plasma Lane RS, Rock GA, Smit Sibinga CTh. (eds). Proceedings from the Netherlands Red Cross FVIII Workshop. Groningen: 1981: 26-30
- 3 Rock GA, Smiley RK, Tittley P, Palmer DS. Factor VIII concentrates: In vivo effectiveness of a high yield product prepared in the blood bank. N Engl J Med 1984; 311: 310-313
- 4 Rock GA, Cruickshank WH, Tackaberry ET, Ganz PR, Palmer DS. Stability of VIIIC in plasma: the dependence on protease activity and calcium. Thromb Res 1983; 29: 521-535
- 5 Smith JK. The assay of factor VIII in heparinized blood. In Development systems for efficient separation of Factor VIII from human fresh frozen plasma Lane RS, Rock GA, Smit Sibinga CTh. (eds). Proceedings from the Netherlands Red Cross FVIII Workshop. Groningen: 1981: 23-25
- 6 Kemball-Cook G, Furling RA, Peake IR, Barrowcliffe TW. Aluminum hydroxide adsorption of factor VIII procoagulant antigen from plasma and factor VIII concentrate. Thromb Res 1981; 23: 193-196
- 7 Breckenridge RT, Ratnoff OD. Studies on the nature of the circulating anticoagulant directed against antihemophilic factor: With notes on an assay for antihemophilic factor. Blood 1962; 20: 137-147
- 8 Pool JG, Robinson J. Assay of plasma antihaemophilic globulin (AHG). Br J Hematol 1959; 5: 17-30
- 9 Poon MC, Ratnoff OD. Immunologic evidence that the antihemophilic factor (factor VIII)-like material in hemophilic plasma possesses a nonfunctional low molecular weight subcomponent. Blood 1977; 50: 367-376
- 10 Strauss HS, Merler E. Characterization and properties of an inhibitor of Factor VIII in the plasma of patients with hemophilia A following repeated transfusion. Blood 1967; 30: 137-150
- 11 Zimmerman TS, de la Pointe L, Edgington TS. Interaction of factor VIII antigen in hemophilic plasmas with human antibodies to Factor VIII. J Clin Invest 1977; 59: 984-989
- 12 MacFarlane DE, Stibbe J, Kirby EP, Zucker MB, Grant RA, McPherson J. A method for assaying von Willebrand factor (ristocetin cofactor). Thromb Diathes Haemorrh 1975; 34: 306-307
- 13 Rick ME, Hoyer LW. Molecular weight of human factor VIII procoagulant activity. Thromb Res 1975; 7: 909-916
- 14 Rock GA, Palmer DS, Tackaberry ET, Cruickshank WH. The presence of high and low molecular weight forms of factor VIII in heparinized plasma. Thromb Res 1978; 13: 85-96
- 15 Rock GA, Tackaberry ET, Palmer DS. Factors affecting the relative distribution of high and low molecular weight forms of factor VIII. Thromb Res 1979; 14: 573-587
- 16 Kirkwood TB L, Rizza CR, Snape TH, Rhymes IL, Austen DE G. Identification of sources of inter-laboratory variation in factor VIII assay. Br J Haematol 1979; 37: 559-568
- 17 Lang H. Aluminum hydroxide adsorption and factor VIII clotting assays. Lancet 1980; 2: 154
- 18 Seghatchian MJ, Kemball-Cook G, Barrowcliffe TW. Adsorption of factor VIII by aluminum hydroxide. Haemostasis 1979; 8: 106-116
- 19 Rock G, Palmer DS. Accumulative effect of DDAVP and heparin in increasing plasma factor VIII levels. Vox Sang 1981; 41: 56-60
- 20 Smit Sibinga CTh, Welbergen H, Das PC. High yield method of production of freeze-dried purified Factor VIII by blood banks. Lancet 1981; 2: 449-450
- 21 Smit Sibinga CTh, Daenen SM G J, van Imhoff GW, Maas A, Das PC. Heparin small pool high yield purified Factor VIII: In vivo recovery and half-life of routinely produced freeze-dried concentrate. Thromb Haemostas 1984; 51: 12-15
- 22 Krachmalnicoff A, Thomas DP. The stability of Factor VIII in heparinized plasma. Thromb Haemostas 1983; 49: 224-227