I'd have to list GlaDOS, Teyrn Loghain and Frank Fontaine up there for me too. I also really like the Prophets from Halo (I can't resist a good religious fanatic), the Dark Presence from Alan Wake and Regnier from Kingom Under Fire.
Most BioWare villains are pretty great. Saren, Darth Malak, the Spoiler Bad Guy from Jade Empire.
However, I would say that Nergal from Fire Emblem is my favorite villain, not only is he incredibly evil, but he actually does stuff. In a game where characters die permanently only due to your own mistakes, him killing off characters is just a hard thing to take.
Everyone here has listed my favorites (Ocelot being probably at the top) but after recently replaying InFamous, I really like Kessler. In my opinion he was a great antagonist.
Lionel Starkweather. One of my favourite celebrity performances in gaming. Brian Cox was unfucking believable, and was as much part of the game as were the executions (and was probably just as disturbing). Also on the topic of Manhunt I imagine Piggsy would make my list if I were to make one too. Piggsy and Pyramid Head should star in a sitcom together.
I can't say wich is the best. One of the most memorable villian, for sure, is The Director from the first Manhunt. That voice just... yeah... it made an already distrubed game even more so.
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