Tudyk (pronounced Tude-ick) was born in El Paso, Texas, the son of Betty Loyce (née Wiley) and Timothy Nicholas Tudyk. His father is of Polish descent and his mother has German, English, and Scottish ancestry. He was raised in Plano, Texas, a suburb of Dallas, where he attended Plano Senior High School. He had a brief experience as a stand-up comic, before quitting after an angry audience member threatened to kill him. Tudyk studied drama at the Methodist-affiliated Lon Morris College in Jacksonville, Texas where he won the Academic Excellence award for drama. While in college, he played Beaver Smith in an eastern New Mexico summer stock theater production of Billy the Kid. Tudyk entered Juilliard but left in 1996 before earning a degree.
In 2000, Tudyk played Gerhardt, a gay German drug addict, alongside Sandra Bullock and Viggo Mortensen in 28 Days.
Tudyk played Wat in 2001's A Knight's Tale, as well as 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, he 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 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. Douglas. Tudyk also voiced the character King Candy in the 2012 Disney animated film, Wreck-It Ralph, a performance for which he won the Annie Award for Voice Acting. He plays Ben Chapman in the 2013 movie 42 about Jackie Robinson.
Among Tudyk's better known roles was the playful, easy-going Hoban "Wash" Washburne in the television series Firefly by Joss Whedon. Although the series ran for only one season, Universal Studios bought the rights to the show and turned it into a film, Serenity, in which he reprised the role. He did a few voices from Make Way for Noddy.
Tudyk appeared as the cult leader, Father, in a two-part episode of Strangers With Candy entitled "Blank Stare." Among several guest spots on popular shows such as Arrested Development, he played a convicted pedophile on a popular episode of CSI: Crime Scene Investigation. Tudyk was cast as a special guest star in Joss Whedon's Dollhouse. The show featured people whose personalities had been erased, with Tudyk portraying Alpha, a former "active" who accidentally downloaded 48 separate personalities. Alpha served as the main antagonist of the series' first season, with guest appearances in the show's second season. Tudyk also guest starred in three episodes of ABC's modern remake of the television miniseries V. Tudyk provided the voice of superhero Green Arrow and villain Psimon in the animated series Young Justice. He is co-starring in the ABC comedy series Suburgatory as Noah Werner, a dentist from the big city who moved to the suburbs.