Molecular design, binding, and APC-generating activities of scFv/TM. (A) Schematic diagram describing the cloning strategy for the fusion construct scFv/TM. The final construct contains a triple FLAG tag at the C-terminus introduced for purification (not shown). (B) Binding of ¹²⁵I-scFv/TM to mouse RBCs after 1-hour incubation with a 1% suspension of washed RBCs and removal of free protein (precoating). Open circle in bottom right corner represents binding to human RBCs to control for specificity. (C) In vitro generation of APC by thrombin is stimulated by mouse RBCs preloaded with scFv/TM. Human RBCs preloaded with scFv/TM and unloaded mouse RBCs serve as negative controls. Unless indicated otherwise, data in this and figures that follow are shown as mean ± SEM (n = 3). Please note that deviation bars are often too small to be evident.