After earning his M.F.A., Jeffrey Donovan made his film debut in Inside Out. His next film, Throwing Down, received the top jury prize at The Hamptons International Film Festival. Donovan also portrayed Jeffrey Patterson in Book of Shadows: Blair Witch 2 and opposite Kevin Bacon in Barry Levinson's Sleepers. In 2005 Jeffrey Donovan appeared as a minor character in the romantic comedy Hitch. The following year he was cast as Cal in Come Early Morning. and as the coach of a women's basketball team in the film Believe in Me. In Changeling, directed by Clint Eastwood and released late 2008, Donovan starred opposite Angelina Jolie as J.J. Jones. Jeffrey was also in When Trumpets Fade.

Donovan has appeared in CSI: Miami, Spin City, Monk, Witchblade, The Pretender, Homicide: Life on the Street, and Law & Order. In the short-lived TV series Threshold, Donovan portrayed the antihero Dr. Sloan. In Crossing Jordan he was cast in the recurring role of William Ivers.

In 2004 he received his first leading role in the American remake of the British television series, Touching Evil opposite Vera Farmiga on the USA Network. Since 2007 he has starred as the lead in Burn Notice. On Burn Notice Donovan plays Michael Westen, a spy who was burned, cast away, and now is trying to return to his prior spy career. Heading into its 4th Season on the USA network, Burn Notice is the most popular program on cable and consistently beats the network competition.

Donovan made his directorial debut directing the Burn Notice episode "Made Man," which first aired on June 17th, 2010.

Donovan has also performed on the stage including the eponymous role Hamlet, A View from the Bridge as Marco, An Inspector Calls and Off-Broadway in Things You Shouldn't Say Past Midnight as Gene, The Glory of Living as Clint, regionally in Toys in the Attic as Julian Berniers, On the Waterfront as Terry, Oedipus as Teiresias and Freedomland as Seth. From October 2008 until spring 2009, Jeffrey starred in the comedy farce Don't Dress for Dinner in Chicago. Also performances on the radio: On the Waterfront as Terry, Frozen and Grapes of Wrath.

Donovan was born in Amesbury, Massachusetts, and raised, along with his two brothers, by their mother. Of his childhood, Donovan recalled:

At Amesbury High School, he was mentored by teacher Patty Hoyt. She aided him in starting a drama club and then found him a private scholarship, enabling him "to participate in a summer program that helped pave the way for his acting career." Donovan went to college at Bridgewater State College, then transferred to and graduated from the University of Massachusetts Amherst. He then went on to earn an M.F.A. degree from New York University’s Tisch School of the Arts.

In California, Donovan owns a home in a rural area overlooking the 11,000-acre Topanga State Park. He resides in Florida where Burn Notice is filmed six months of the year.

Donovan is also a black belt in karate, has studied aikido for about six years, and Brazilian jiu-jitsu for about a year.