In Vivo Recovery of Factor VIII: A Comparison of One-Stage and Two-Stage Assay Methods

Summary

The recovery and half-life of VIII: C in the plasma of severely haemophilic patients was measured by one-stage and two-stage assays after injection of two Factor VIII concentrates (Hemofil, Hyland and Fraction I-O, Kabi). Plasma volumes were measured with an Evans� Blue technique, and both concentrates and post-infusion samples were measured against the same plasma standard.

There was a highly significant difference in recoveries estimated by the two assay methods. The one-stage assays gave the most consistent results, in that the average recovery was 100%, whereas the two-stage assays gave only about 80% of the value expected from in vitro assays. There was no difference in recoveries between the two concentrates.

The two-stage assays gave a slightly shorter half-life than the one-stage assays, and the half-life of Hemofil was also shorter than that of Fraction I-O.

• References

• 1 Adelson E. 1964. The survival of factor VIII and factor IX in normal humans. In Brinkhous KM. (Ed.) The Haemophilias. p 197 University of North Carolina Press; Chapel Hill:
• 2 Biggs R. 1964; The turnover of factors II, VIII, IX and X in patients deficient in these factors. Thrombosis et Diathesis Haemorrhagica Suppl 13: 201
• 3 Biggs R, Matthews JM. 1966. The plasma concentration of factor VIII in the treatment of haemophilia. In Biggs R, Macfarlane RG. (Eds.) Treatment of Haemophilia and Other Coagulation Disorders. Blackwell; Oxford: p 107
• 4 Biggs R, Eveling J, Richards G. 1955; The assay of antihaemophilic globulin activity. British Journal of Haematology 1: 20
• 5 Blombäck B, Blombäck M. 1958; Purification of antihaemophilic globulin. I. Some studies on the stability of the antihaemophilic globulin activity in fraction 1-0 and a method for its partial separation from fibrinogen. Arkiv för Kemi 12: 387
• 6 Johnson AJ, Karpatkin MH, Newman J. 1971; Clinical investigation of intermediate- and high-purity antihaemophilic factor (factor VIII) concentrates. British Journal of Haematology 21: 21
• 7 Kirkwood TB L, Rizza CR, Snape TJ, Rhymes I, Austen DE G. 1977; Identification of sources of inter-laboratory variation in factor VIII assay. British Journal of Haematology 37: 559
• 8 Kirkwood TB L, Barrowcliffe TW. 1978; Discrepancy between one-stage and two-stage assay of Factor VIII :C. British Journal of Haematology 40: 333
• 9 Laurell CB. 1965; Antigen-antibody crossed electrophoresis. Analytic Biochemistry 10: 358
• 10 Nilsson I-M, Blomback M, Von Francken I. 1957; On an inherited autosomal hemorrhagic diathesis with antihemophilic globulin deficiency and prolonged bleeding time. Acta Medica Scandinavica 159: 35
• 11 Nilsson I-M, Blomback M, Ramgren O. 1962; Haemophilia in Sweden: VI. reatment of haemophilia with the human antihaemophilic factor preparation (fraction I-O). TActa Medica Scandinavica Suppl 379: 61
• 12 Nilsson I-M. 1974. Haemorrhagic and Thrombotic Diseases. John Wiley and Sons; London:
• 13 Nilsson I-M, Hedner U, Ahlberg Å, Larsson SA, Bergentz S-E. 1977; Surgery of haemophiliacs - 20 years’ experience. World Journal of Surgery 1: 55
• 14 Nilsson I-M, Hedner U. 1977; Characteristics of various factor VIII concentrates used in treatment of haemophilia A. British Journal of Haematology 37: 543
• 15 Nilsson I-M, Barrowcliffe TW, Kirkwood TB L, Miller-Andersson M, Seghatchian MJ. 1977; Problems of quantitation of FVIII:C activity. 6th Congress of the International Society for Thrombosis and Haemostasis (Abstract) Thrombosis and Haemostasis 38: 25
• 16 Over J, Sixma JJ, Poucet-De Bruine MH M, Trieschaigg AM C, Vlooswuk RA A, Beeser-Visser NH, Bouma BN. 1978; Survival of ¹²⁵Iodine-labelled factor VIII in normals and patients with classic haemophilia. Journal of Clinical Investigation 62: 223
• 17 Owen Jr C, Bowie EJ W. 1975. Infusion therapy in hemophilia A and B. In Brinkhous KM, Hemker HC. (Ed.) Handbook of Hemophilia. Part II. p 487 Excerpta Medica; Amsterdam:
• 18 Peake IR, Bloom AL. 1978; Immunoradiometric assay of procoagulant factor VIII antigen in plasma and serum and its reduction in haemophilia. Lancet 1: 473
• 19 Pool JG, Robinson J. 1959; Observations on plasma banking and transfusion procedures for haemophilic patients using a quantitative assay for antihaemophilic globulin (AHG). British Journal of Haematology 5: 24
• 20 Reitzlaff JA, Tauxe WN, Kiely JM, Stroebel CF. 1969; Erythrocyte volume, plasma volume and lean body mass in adult mean and Women. Blood 33: 649
• 21 Smith CM, Miller GE, Breckenridge RT. 1972; Factor Vffl concentrates in outpatient therapy. Journal of the American Medical Association 220: 1352
• 22 Wagner RH, Mclester WD, Smith M, Brinkhous KM. 1964; Purification of antihemophilic factor (factor VIII) by aminoacid precipitation. Thrombosis et Diathesis Haemor- rhagica 11: 64