Titers of FVIII-binding antibodies assessed for individual Ig isotypes and IgG subclasses. Presented are the results for FVIII-binding antibodies for all plasma samples. The screening cutoff of the methods to differentiate positive from negative samples is at a titer of 1:20. The lines at a titer of 1:40 respectively 1:80 represent the lower limit of quantification (LLOQ) for confirmed FVIII specificity of the different Ig isotype/IgG subclass antibodies based on successful FVIII-competition.31,32 Circles depict positive results for FVIII-binding antibodies with confirmed FVIII specificity; triangles represent results below the FVIII-specific LLOQ. (A) Thirty-three of 81 patients with nsHA, (B) 10 of 39 patients with sHA, and (C) 11 of 90 healthy donors contained FVIII-binding antibodies with confirmed FVIII specificity. Titers highlighted in red belong to patient 58 (nsHA). FVIII, factor VIII; Ig, immunoglobulin; ND, not detectable.