Figure 2.

Manufacture and characterization of t-TLNPs. (A) Bioconjugation schematics of reacting cysteine-terminated peptides to maleimide-terminated DSPE-PEG_2K utilizing thiol–maleimide chemistry to synthesize DSPE-PEG_2K-peptide molecules. (B) Molecular components of t-TLNP manufacture. (C) Dynamic light scattering (DLS) analysis of five representative t-TLNP batches showing nanoparticle size reproducibility. (D) Cryo-transmission electron microscopy (Cryo-TEM) images of t-TLNPs (scale bar: 100 nm) showing a particle diameter of ∼175 nm. (E) Representative images from BioFlux experiments where Rhodamine B labeled control (undecorated) vs targeted nanoparticles (“VBP + CBP”-decorated) were flowed at 25 dyn/cm² over “vWF + collagen”-coated channels and targeted nanoparticles were also flowed over albumin-coated channels, showing substantially high adhesion of targeted nanoparticles to “vWF + collagen”-coated surface but not of control particles to the “vWF + collagen”-coated surface or targeted nanoparticles to the albumin-coated surface. (F) Spectrometric analysis of three representative t-TLNP batches showing a mean thrombin loading of 114.3 ±14.2 nM. (G) Thrombin release analysis showing that t-TLNPs can slowly release low amounts of thrombin by diffusion, whereas exposure to sPLA₂ significantly enhances thrombin release. *p≤ 0.05, **p≤ 0.01.