Studies on the Adsorption of Factor VIII: Application to the Purification of Bovine Factor VIII

Simonetti, C; G, Casillas; Pavlovsky, A

doi:10.1111/j.1365-2141.1972.tb03456.x

Cited by 6 publications

(2 citation statements)

References 8 publications

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“…The results of gel filtration studies may be difficult to interpret, because they may be influenced so easily by the experimental conditions. It is known, for example, that a change in the molarity or the pH of the eluant may alter the elution pattern of factor VIII (Owen & Wagner, 1972;Weiss et al, 1972), and factor-VIII clotting activity, particularly when it is separated from other plasma proteins, may be inactivated by trisodium citrate (Simonetti et al, 1972). The results of our experiments using different eluants reflect this instability of factor-VIII clotting activity.…”

Section: Discussionmentioning

confidence: 53%

Transfusion and Gel Filtration Studies in von Willebrand's Disease

1974

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A study has been made of the plasma levels of factor-VIII clotting activity and factor-VIII related antigen following transfusion in five patients with von Willebrand's disease and one patient with severe haemophilia. All ofthe patients were transfused with cryoprecipitate or an AHG concentrate and their plasma levels of factor-VIII clotting activity and factor-VIII related antigen assayed at intervals after transfusion. In the case of the patients with von Willebrand's disease there was the expected post transfusion rise of factor-VIII activity followed by a fall of activity and then a secondary rise of activity. During this same period there was a rapid fall

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Section: Discussionmentioning

confidence: 53%

Transfusion and Gel Filtration Studies in von Willebrand's Disease

1974

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“…[5], Preparation of rabbit antibodies: highly purified, EDTA-inactivated, thrombin-treated bovine factors VIII and bovine factor VIII inhibitor complex were used as antigens. Of these materials, 2 ml containing 2 mg were administered intramuscularly with Freund's complete adjuvant (2 ml) with 2 weeks interval.…”

Section: Methodsmentioning

confidence: 99%

Physical, Chemical and Immunological Studies on Bovine Factor VIII

G¹,

Simonetti²,

Vásquez³

et al. 1976

Pathophysiol Haemos Thromb

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Highly purified bovine factor VIII prepared according to our technique and having a specific activity of 500 U/mg protein has been studied. The chemical analysis of the preparation revealed it to be composed of amino acids, lipids (8–10%) and carbohydrates (7 %). The lipid moiety can be removed by chromatography. Different mechanisms of inactivation of bovine factor VIII and the possible molecular changes induced by the inactivating agents were studied. EDTA and EGTA provoke a weakening of the bonds linking the structural elements of the molecule, which allows for the separation of two different components of the molecule by gel filtration. Upon treatment with thrombin, the carbohydrate content of bovine factor VIII decreases without any apparent degradation of the protein moiety of the molecule. The fact that precipitating and neutralizing antisera against factor VIII were obtained, shows that the molecules modified by EDTA and thrombin still have the antigenic properties of factor VIII. The inhibitor developed by hemophiliacs transfused with human factor VIII is bound to bovine factor VIII, forming a complex which is stable upon agarose gel filtration. Immunological studies of the purified complex reveal that it is composed of bovine factor VIII and human γ-globulin. Bovine factor VIII in the complex retains some antigenic determinants which bind rabbit anti-serum against bovine factor VIII, as shown by neutralization of the antiserum and by precipitation studies.

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