This game has twenty-two characters taken from both the worlds of DC Comics and Teenage Mutant Ninja Turtles. As well as the four famous turtles, this game also has other well-known characters from their universe such as April O'neil and Shredder. On the other hand, there are also popular characters from the DC universe, such as Batman, Superman, Wonder Woman, and even some villains like DarkSeid and Despero.
Although this game has some original sprites and soundtracks, most of its content is taken from two popular 16-bit games: TNMT Tournament Fighters and Justice League Task Force. Although those games come from two different studios (Konami and Acclaim) the color palette has been mortified and integrated with the scenes to perfectly combine the two universes.
As well as a classic arcade game mode, Teenage Mutant Ninja Turtles vs Justice League also has a local multiplayer mode, a training mode, and a special game mode in which you play as a random character.