Spider-Man takes Mary Jane Watson, his girlfriend, on a thrill ride, and after that, talks about Harry. A small meteor crashes, with an alien symbiote inside. Meanwhile, an escaped convict Flint Marko falls into a lab pit and turns into sand. Peter's former best friend Harry Osborn does not talk to him anymore and he has a new rival in the Daily Bugle named Eddie Brock. He defeats the bombers of the building and saves a woman from being killed. The game starts off as Spider-Man swings into a burning building.

Hulk (Bruce Banner) (Mentioned in out-of-universe commentary).

Andrews (First and only known appearance) (Xbox 360, PS3 and PC versions only) Chen (First and only known appearance) (Xbox 360, PS3 and PC versions only) Stillwell (First and only known appearance) (Xbox 360, PS3 and PC versions only)

Dragon Tail Gang (First and only known appearance).Arsenic Candy Gang (First and only known appearance).Apocalypse Gang (First and only known appearance).Electro (Max Dillon) (First and only known appearance) (GBA version only) (Mentioned in Xbox 360, PS3 and PC versions).

Shriek (Frances Barrison) (First and only known appearance) (PS2, PSP and Wii versions only).Michael Morbius (First and only known appearance) (PS2, PSP and Wii versions only).Scorpion (MacDonald Gargan) (Last appearance) (Xbox 360, PS3 and PC versions only).Kraven The Hunter (Sergei Kravinoff) (Last appearance).Kingpin (Wilson Fisk) (First and only known appearance dies) (Xbox 360, PS3, PC and GBA versions only).Mad Bomber (Luke Carlyle) (First and only known appearance).Calypso (Last appearance) (Xbox 360, PS3 and PC versions only).Curt Connors) (First appearance as Lizard) Rhino (Alexander O'Hirn) (Last appearance) (Xbox 360, PS3 and PC versions only).

Captain Jean DeWolfe (First and only known appearance) (Xbox 360, PS3 and PC versions only).

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