Construction and characterization of PLT biomimetic N+R@PLTs

PLTs (1 × 109) were resuspended in 1 ml of PBS, incubated with a 1:100 dilution of a biotinylated anti-CD42a monoclonal antibody for 30 min, and then washed with PBS. Afterward, PLTs were incubated with avidin solution (2 mg/ml) for another 30 min and then washed again. Last, PLTs were incubated with 100 μg of photothermal nanoparticles (which had been linked to biotin) in 1 ml of PBS for another 30 min and then washed to obtain N@PLTs. During the incubation, an adjuvant imiquimod hydrochloride (R) solution was added into the medium in a final concentration of 80 μM to obtain N+R@PLTs. The final loading efficiencies of N and R were about 4.4 and 10.2%, respectively. The concentration of N+R@PLTs in photothermal effect study in vitro was 1 × 1010 N+R@PLTs/ml (~43.8 μg/ml or ~44.5 μM for N). For animal experiments, the doses of PLTs, N, and R were 1 × 109 cells per mouse, ~4.4 μg per mouse, and 2.0 μg per mouse, respectively. In SIM image, R was replaced by a blue-emitting dye, coumarin derivative. The size and surface potential of N, PLTs, and N+R@PLTs were detected by dynamic light scattering (Malvern ZEN 3600 Zetasizer, UK).

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