Evaluation of LIBS exposure on the patient's platelets.
To assess spontaneous LIBS exposure on platelets, PRP (2 × 106 platelets) with 10−7 M PGE1 was incubated with 10 μg/mL anti-LIBS antibodies 7D6, AP5, and D3 in the presence of 2 mM RGDS or RGES peptides for 60 minutes at RT. After 2 washes, platelets were incubated with FITC-labeled goat anti–mouse F(ab′)2 for 30 minutes, then samples were washed once and bound antibodies were analyzed by flow cytometry. LIBS antibody binding was expressed as a LIBS index (LI) by standardizing the MFI for each LIBS monoclonal antibody (mAb) to that obtained using AP3, the binding of which was not affected by the patient's mutation. LI = LIBS mAb MFI/AP3 MFI for the patient and for the control separately.
