Of particular note is that the title is misleading – the plot of the comic book series Venom: Separation Anxiety involves Venom's five symbiote spawn attempting to force Eddie (without his symbiote) to help them communicate with their own symbiotes. The events of the game more closely follow the events of Venom's first limited series, Venom: Lethal Protector.
Spider-Man – The titular hero of the series, Spider-Man is weaker and faster than his teammate, Venom. As in the comic, he is able to punch, kick, swing from webs and cling to walls. In combat, his special moves are oriented toward agility and finesse rather than brute strength.
Venom – As a symbiote, Venom is a combination of both human and alien organism. He has all of Spider-Man's generic abilities (web-swinging and wall-crawling), but unlike Spider-Man, his combat moves are primarily based on raw power. He also takes less damage per hit than Spider-Man.
Carnage- The main antagonist of the game, and appears as the final boss.
The Jury – Unlike the Symbiotes, these level bosses are not superhuman; instead, they are extremely well armed and well equipped mercenaries. They appear singly and in pairs at the end of selected levels, and again together at the end of the game. The members are Sentry, Ramshot, Screech, Bomblast, Firearm and Wysper.
Symbiotes – Like the Jury, these hybrid alien/human level bosses possess varied combat skills and attack either alone or in pairs until the end of the game. The symbiotes are Riot, Agony, Lasher, Scream and Phage.