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As much as I'd love to mention an old-school boss like the Tyrant or Nemesis, I have to be honest and go with one of the bosses from RE4. There are just too many memorable moments with the bosses in that game to ignore and I think my favourite would have to be Del Lago.
You can just feel the intensity of that fight, the way you're frantically trying to maneuver the boat while throwing harpoons at the thing. The way the water splashes at the screen, and Leon's desperate swim for the boat just makes the experience all the more visceral.
I just stole 2 seconds from your life. No, you can't have them back...