Polystyrene derivatives as candidates for extracorporeal adsorption of Factor VIII antibodies in the management of haemophiliac patients.

BACKGROUND AND OBJECTIVES: This article presents a new approach for removing the Factor VIII inhibitors (anti-FVIII) in haemophiliac patients by immunoadsorption using an affinity matrix. MATERIALS AND METHODS: Ten blood samples from haemophiliac patients with anti-FVIII were assayed for antibodies, total immunoglobulins, procoagulant proteins and complement C3 protein after circulation over one or two columns filled with the polymers under investigation. RESULTS: These new synthetic sorbents are able to remove in vitro 90% of anti-FVIII from haemophiliac plasma with inhibitors (up to 540 Bethesda Units/ml). Neither coagulation factor adsorption nor effects on complement system activation were observed. CONCLUSIONS: The data presented clearly show that these polymers allow a rapid and efficient reduction of inhibitor titre. In view of the parameters studied, these polymers fulfil the requirements for use in a blood purification process to decrease high inhibitor titres without losing essential proteins.

Adsorption of immunoglobulin G and anti-factor VIII inhibitory antibody from haemophiliac plasma to derivatized polystyrenes.

Modified polystyrene resins containing sulphonate groups and tyrosyl sulphamide or tyrosyl methyl ester sulphamide groups have been investigated with respect to their potential for selective binding of anti-Factor VIII inhibitory antibodies from plasma. Adsorption of total immunoglobulin G and of a monoclonal antibody to Factor VIII was measured following addition of the radioiodinated proteins to normal plasma, plasma depleted of Factor VIII by adsorption on a resin coupled to anti-Factor VIII antibody, and haemophiliac plasma containing various levels of inhibitory anti-Factor VIII antibody. Depletion of anti-Factor VIII antibody from the haemophiliac plasmas by incubation with the resins was also measured by Bethesda assay. The modified resins and their corresponding unmodified "controls' showed similar binding of total immunoglobulin G. However, only resins containing either sulphonate or a combination of sulphonate and tyrosyl sulphamide groups showed evidence of selective adsorption of anti-Factor VIII antibody from plasma.

Affinity of human anti-factor VIII antibodies for functional polystyrene supports.

Human anti-factor VIII antibodies (anti-FVIII) neutralize Factor VIII (FVIII) procoagulant activity. These antibodies appear in about 5-15 per cent of severely affected patients with haemophilia A treated with FVIII concentrates (Mannucci, 1993). In order to obtain non-thrombogenic materials able to interact specifically with anti-FVIII, amino acids residues that mimic part of the FVIII molecule recognized by anti-FVIII have been grafted. Several cross-linked polystyrenes were functionalized with sulphonate and tyrosine sulphamide groups or tyrosine derivatives sulphamide groups such as methyl ester tyrosine, or the peptides aspartic acid methyl amide tyrosine, tyrosine aspatic acid methyl amide or aspartic acid aspatic acid methyl amide tyrosine. The in vitro removal of anti-FVIII from haemophilic A plasma was performed on different supports. These polymers exhibit strong and selective affinity for the anti-FVIII. The amount of adsorbed anti-FVIII varies with the composition of the polymer and a maximum is achieved for 15-35 per cent of amino acid sulphamide groups. The influence of different chemical groups on the surface of the polymeric solid supports on the adsorption of anti-FVIII was also studied.

Synthetic sorbents for removal of factor VIII inhibitors from haemophilic A plasma.

Human factor VIII (FVIII) is a protein of the blood coagulation system that is absent or defective in patients with haemophilia A. A most serious complication following replacement therapy in 10-15% of patients treated with available FVIII concentrate is the development of inhibitors of FVIII (anti-FVIII). Some polymers functionalized with suitable chemical substituents which mimic part of the epitope of FVIII recognized by the inhibitors might be used in extracorporeal circulation to reduce the concentration of antibodies to FVIII. For this purpose, insoluble polystyrene bearing sulfonate and L-tyrosine methyl ester sulfamide groups have been synthesized. The in vitro removal of anti-FVIII inhibitors from plasmas of patients with haemophilia A was performed. Different chromatographic parameters were studied and optimized.

Tyrosyl sulfamide derivatives as ligands for affinity adsorption of anti FVIII antibodies.

Human anti factor VIII (anti FVIII) antibodies neutralize factor VIII procoagulant activity. These antibodies appear in about 5-10% of severely affected haemophiliac A patients treated with FVIII concentrates. In order to obtain non-thrombogenic materials able to interact specifically with anti FVIII, we have grafted amino acid residues which mimic part of the epitope of the FVIII molecule recognized by the anti FVIII. For this purpose, crosslinked polystyrenes functionalized with sulfonate and tyrosyl or methyl ester tyrosyl sulfamide groups have been synthesized and characterized. The in vitro removal of anti FVIII from haemophiliac patient plasma with antibodies, was performed on these different active supports. These polymers exhibit strong and selective affinity for the anti FVIII. The adsorption of the antibodies vary with the percentage of units bearing methyl ester tyrosyl sulfamide groups and present a maximum at 25% grafting rate. For the more efficient resin, the affinity constant, determined for the adsorption isotherm for the anti FVIII is about 10(9) M-1, whereas the affinity constant for the IgG in the same experimental conditions, is low (10(5) M-1). The influence of different chemical groups on the polymeric phase, on their affinity for the inhibitors was also studied. The most active resins can be selected and used in an extracorporeal circulation to reduce the concentration of anti FVIII in a blood epuration process.