Herein,　we　integrate　the　Hepa1-6　liver　cancer-specific　neoantigen,　toll-like　receptor　9　agonist　and　stimulator　of　interferon　genes　agonist　by　silk-hydrogel　package,　and　combine　with　TIM-3　blockade　to　elicit　robust　antitumor　immunity　for　effectively　suppressing　orthotopic　hepatocellular　carcinoma　(HCC)　progression.　Unlike　intradermal　injection　of　simple　mixed　components　with　short-term　immune　protection,　the　neoantigen　immunotherapeutic-gels　evoke　long-term　immune　protection　to　achieve　significant　prophylactic　and　therapeutic　activity　against　HCC　through　only　one-shot　administration　without　any　side　effects.　Notably,　the　synergized　immunotherapy　by　further　combining　NGC-gels　with　TIM-3　antibody　significantly　reduces　regulatory　T-cells　and　increases　the　IFN-γ　and　IL-12p70　levels　in　tumor　tissues　for　promoting　the　infiltration　of　IFN-γ+CD8+T-cells　and　41BB+CD8+T-cells　to　achieve　complete　remission　(4/7)　and　prevent　pulmonary　metastasis　in　orthotopic　HCC,　and　establish　long-term　memory　against　tumor　rechallenge　with　remarkably　longer　survival　time　(180　days).　Overall,　this　study　provides　an　attractive　and　promising　synergistic　strategy　for　HCC　immunotherapy　with　possible　clinical　translation　prospects.　©　2022　American　Chemical　Society