Penelope is a teenage humanoid girl with tanned skin, brown waist-length hair, and green eyes. She wears a green forest-themed dress, a crown of twigs and leaves, and a black choker with a green spider charm. True to her name, Princess Spiderbite also has a large spider bite on the right side of her forehead - up until it is healed by Slime Monster in "Monster Bash".
Princess Spiderbite first appears in "Club Snubbed" as the representative of the royal Spiderbite family during the annual Silver Bell Ball. She has one word of dialogue in the episode, saying hello to Star Butterfly, and she appears sharing dances with Star, Tom Lucitor, and Princess Jaggs.
In "Monster Bash", Penelope attends Star's party at the Monster Temple to unite Mewmans and monsters. She meets Slime Monster, and they become friends after he uses his slime to heal her spider bite. When the two accidentally stumble into a secret temple chamber, Mina Loveberry attacks them and captures Slime, and Penelope gets blamed by the other monsters for Slime's disappearance.
When Penelope first meets Slime, she is initially wary of him and disgusted by his slimy body. After he heals her spider bite, however, she calls him amazing, and the two become fast friends. She also shows concern for his well-being when they stumble deep into the temple and she squishes his body on the floor.
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- Princess Spiderbite's first name is revealed in her signature on Star Butterfly's "Petition of Friendship" in "Death Peck".