Figure 3. Humanized nano pro-resolving medicines (NPRMs): Construction and characterization.
(A) Human PMN-derived MPs (red) were isolated and enriched with AT-RvD1 or o-[9,12]-benzo-ω6-epi-LXA4 and fluorescent 1,2-dioleoyl-glycero-3-phospho-L-serine-N (7-nitro-2-1,3-benzoxadiazo-4-yl). Enriched NPRMs were separated by size-exclusion chromatography and sized by flow cytometry (blue). (B) Fluorescence incorporation into NPRM (blue histogram) was monitored by flow cytometry. (C & D) NPRM sizing was determined using cytometry calibration beads. (E & F) Sizing was also validated using electron microscopy of MPs and NPRMs following negative staining, calibration nanospheres are shown inset. (G) Incorporation of AT-RvD1 into NPs was determined using LC-MS/MS with representative mass spectra.