Here is a list of villains that appear in Gotham Knights:

• Mr. Freeze
• Penguin
• Clayface
• Harley Quinn
• The Court of Owls
• Ra’s al Ghul
• Talia al Ghul
• Professor Pyg

These villains are scattered across the Batman Universe and are encountered by the four playable characters in the game: Nightwing, Batgirl, Robin, and Red Hood. The game features the biggest open world of any Batman game to date, with several enemy gangs to defeat and a Day/Night cycle that allows the heroes to plan out their moves before striking out once the sun goes down.