Selective human factor VIII activity measurement after analytical in-line purification.

Autor: Engelmaier A; Analytical Development, Pharmaceutical Science Baxalta Innovations GmbH, Part of Takeda Vienna Austria., Schrenk G; Analytical Development, Pharmaceutical Science Baxalta Innovations GmbH, Part of Takeda Vienna Austria., Billwein M; Analytical Development, Pharmaceutical Science Baxalta Innovations GmbH, Part of Takeda Vienna Austria., Gritsch H; Analytical Development, Pharmaceutical Science Baxalta Innovations GmbH, Part of Takeda Vienna Austria., Zlabinger C; Analytical Development, Pharmaceutical Science Baxalta Innovations GmbH, Part of Takeda Vienna Austria., Weber A; R&D Plasma Derived Therapies Baxalta Innovations GmbH, Part of Takeda Vienna Austria.
Jazyk: angličtina
Zdroj: Research and practice in thrombosis and haemostasis [Res Pract Thromb Haemost] 2022 Oct 13; Vol. 6 (7), pp. e12821. Date of Electronic Publication: 2022 Oct 13 (Print Publication: 2022).
DOI: 10.1002/rth2.12821
Abstrakt: Background: It is essential to measure the activity of factor VIII (FVIII) throughout the life cycle of a coagulation FVIII concentrate. Such measurement in nonclinical pharmacokinetic studies is potentially biased by the presence of endogenous nonhuman FVIII, and certain manufacturing process-related additives can also impact the assay performance. Finally, the presence of FVIII activity-mimicking antibodies poses challenges when measuring FVIII in samples. Therefore, we developed an antibody-based chromogenic FVIII assay, which facilitates the selective and sensitive activity measurement of human FVIII in the presence of animal plasma and interfering agents.
Methods: Plate-bound monoclonal anti-FVIII antibody specifically captured human FVIII, which was then measured with a chromogenic activity assay. A human reference plasma preparation was used to construct the calibration curve. Spike recovery was carried out in a citrated cynomolgus monkey plasma-solvent/detergent mixture and in the presence of the bispecific antibody emicizumab.
Results: The calibration curve ranged from 3.03 to 97.0 mIU FVIII/ml and was obtained repeatedly with good accuracy. B domain-deleted and full-length FVIII did not differ in their responses. Recovery of spiked human FVIII in citrated cynomolgus monkey plasma was 102.7%, while neither native monkey plasma nor the other animal specimen tested showed any activity. Solvent/detergent solution and the bispecific antibody emicizumab had no influence on the assay.
Conclusion: Combining antibody-mediated specific capture of human FVIII and a chromogenic activity assay resulted in a selective and sensitive measurement of human FVIII with no interference by endogenous, nonhuman FVIII, manufacturing process additives, or an FVIII activity-mimicking antibody.
(© 2022 The Authors. Research and Practice in Thrombosis and Haemostasis published by Wiley Periodicals LLC on behalf of International Society on Thrombosis and Haemostasis (ISTH).)
Databáze: MEDLINE
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