The game was released as a coin-operated arcade title based on the Sega System 32 hardware. The game can be played as a single player game or up to a four players cooperatively. Each character can do a special move related to their super power which reduces their health.

During levels the game changes from a side-scrolling beat 'em up into a platform game as the camera zooms to a far-away view of the characters in miniature. Later it zooms back in for the much larger and more detailed characters to continue the brawling.

The game was divided into four acts, meeting various villains, including Kingpin, Venom and his symbiote clones, Doctor Octopus, Electro, Lizard, Scorpion, Sandman, Green Goblin, Hobgoblin, and finally Doctor Doom. High scores are separated by character; so a high score on Spider-Man may not be a high score on Hawkeye or Black Cat.

Spider-Man: The Video Game's soundtrack is also partially comprised from tunes from Sega's 1986 arcade game Quartet, notably the tunes "Oki Rap" and "FM Funk" (for instance, "FM Funk" appears in the second stage in Spider-Man, while it is featured in the third stage of Quartet).