Usefulness of chromogenic assays for potency assignment and recovery of plasma-derived FVIII and FIX concentrates or their recombinant long acting therapeutic equivalents with potential application in treated pediatric hemophiliac patients.

On demand and prophylaxis usage of FVIII/ FIX concentrates for the therapeutic management of hemophilia has greatly changed quality of life, and healthy life span of affected patients. Availability of recombinant therapeutic FVIII and FIX products, and of their long-acting variants, further improves the treatment constraints, and progressively permits to hemophiliacs to have an almost normal way of life. Unlimited amounts of recombinant or engineered substitutive products become available, and open new avenues for extending the benefits of prophylaxis to all hemophiliac patients, not only in economically advanced territories, but also in emerging and developing countries, worldwide. Pharmacokinetics of injected products can be variable among treated patients, and dependent on age. In addition, patient medical status, existing diseases, and the nature of joint damages can impact protective effect of substitutive products, and risks associated to way of life and activity. Product requirements and half-life of infused products are therefore patient specific. Monitoring recoveries of injected products thus provide useful information for the most appropriate treatment adjustment. FVIII and FIX measurements in plasma of treated patients helps to establish the optimal interval between injections for each treated patient, and the overall therapeutic cost. Due to the high variability from reagent to reagent, and the different behavior from plasma derived products, clotting methods are not ideal for recombinant and long-acting products. They require to be performed only in association with a drug specific calibrator. They are not recommended for patients' survey, due to the high variety of reagents available. Chromogenic assays (2-stage methods) offer a standard reactivity to all available FVIII or FIX products in drugs, whether the way they are obtained, or in plasma. In a subset of treated patients, inhibitory antibodies to FVIII or FIX develop and can be measured with inhibition assays (Bethesda units), or by Elisa. Unfortunately, FVIII or FIX substitutive therapies cannot be used in patients with inhibitors, and alternative clinical management is requested, such as the use of FEIBA or FVIIa for their bypassing activity. A new treatment is being introduced in the form of a bispecific antibody (Emicizumab) targeted to both FIXa and FX, and which allows activating FX by FIXa without the need for FVIII. Some chromogenic assays (Biophen FVIII), designed with human proteins, offer the possibility to measure the activity and recovery of this new drug. Chromogenic methods are then useful for establishing potency of therapeutic products or monitoring recovery and kinetics in treated patients, through plasma measurements. Availability of International Standards for FVIII and FIX, in concentrates or plasma, allows harmonization of assay results.