|The following is a list of main and recurring characters in the Fresh Beat Band of Spies series.
- Twist is a disc jockey and the show's central character. He is voiced by Jon Beavers.
- Kiki is a guitarist and the lead singer of the band. She is voiced by Yvette Gonzalez-Nacer.
- Shout is an athletic keyboardist. He is voiced by Tommy Hobson.
- Marina is an intelligent drummer. She is voiced by Tara Perry.
- Bo Monkey is Twist's best friend and the four band members' sidekick. He is voiced by Tom Kenny.
- Reed is the gadget guy of the team. He is known for appearing out of nowhere. He is voiced by Tom Kenny.
- Commissioner Goldstar acts as the spies' supervisor and friend. He is voiced by Keith Silverstein.
- Von Winnerchamp is the most frequently-seen antagonist, who loves collecting things. He is voiced by Tom Kenny.
- Lily is Commissioner Goldstar's neice. She is a dog owner and a friend of the Fresh Beats. She is voiced by Heaven White.
- Lil Piggie is a pig with incredible dancing skills. He resides at the pet center.
- Captain Arrrgh is a singing pirate captain who owns a chicken named Poulet. He is voiced by Tom Kenny.
- Poulet is Captain Arrrgh's chicken sidekick. She can lay eggs and clucks often. Poulet is voiced by Keith Silverstein.
- Yi-Haw is a cowgirl and robber, notorious for stealing entire buildings. She is voiced by Kate Higgins.
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