He played Wat in 2001's A Knight's Tale, Steve the Pirate in the 2004 comedy Dodgeball: A True Underdog Story and the emotional robot Sonny in I, Robot. In 2005, he reprised his role as the playful, easy-going Hoban 'Wash' Washburne in the film Serenity, derived from the cult television series Firefly by Joss Whedon. In 2007, Tudyk had a supporting role as a strong-willed doctor in the western film 3:10 to Yuma alongside Christian Bale and Russell Crowe along with a brief but memorable role in the film Knocked Up and a highly physical comedic performance in the British film Death at a Funeral. Tudyk starred as Tucker in the indie horror comedy Tucker & Dale vs Evil. He also appears in Transformers: Dark of the Moon, reprising his role as Dutch.
In Alvin and the Chipmunks: Chipwrecked, he voiced Simone (a reckless French accent speaking Chipmunk, the result after Simon was bitten by a spider). In 2012, Tudyk appeared in Timur Bekmambetov's Abraham Lincoln: Vampire Hunter, in the role of Stephen A. He played Ben Chapman in the 2013 film 42 about Jackie Robinson.
In 2012, Tudyk voiced King Candy in Walt Disney Animation Studios' 2012 film Wreck-It Ralph, a performance for which he won the Annie Award for Voice Acting. As of 2022, Tudyk has lent his voice to every Walt Disney Animation Studios film since.