ArticleThe AAPS journal2026
Challenges in the Quantification of Free TFPI in Human Plasma Following Marstacimab Treatment.
Article in The AAPS journal, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Tissue factor pathway inhibitor (TFPI) is an endogenous inhibitor of the extrinsic coagulation pathway. Marstacimab (PF-06741086), a human immunoglobulin G1 monoclonal antibody, binds the TFPI Kunitz-2 domain to improve hemostatic control in individuals with hemophilia. Quantifying the free form of TFPI, i.e., not bound to marstacimab or endogenous binding proteins, is the most direct assessment of target coverage and modulation following treatment. This investigation describes efforts to develop a quantitative assay to measure free TFPI in human plasma to support the marstacimab drug development program. Three protein enrichment methods (competitive antibody capture, binding partner capture, and drug depletion) were assessed in combination with liquid chromatography-tandem mass spectrometry to develop a bioanalytical free target assay. Surface plasmon resonance (SPR) was used to determine the binding affinities of marstacimab and competitive antibody clones to TFPI. SPR analysis demonstrated marstacimab binds TFPI with a nanomolar affinity (equilibrium dissociation constant [K
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