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The melanin-based vaccine formulations were composed of peptide–melanin nanoaggregates (pOVA30-Mel or mAdpgk-Mel) in which the phosphorothioate oligonucleotide CpG-28 (5′-TAAACGTTATAACGTTATGACGTCAT) (Oligovax, Paris, France), a B-type CpG-ODN [16, 17], was added (50 µg/mouse) just before the immunizations. Control mice were injected with free peptide and 50 μg CpG-28 emulsified in incomplete Freund’s adjuvant (vol/vol) (Sigma-Aldrich, Saint-Quentin-Fallavier, France) or with 50 μg CpG-28 and melanin without peptide. Vaccine formulations were administered subcutaneously in the flank of the mice (100 µl/injection) on the indicated day.
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