Peter Parker / Spider-Man (voice)
Yuri Watanabe (voice)
Wilson Fisk / Additional Voices (voice)
Otto Octavius (voice)
J. Jonah Jameson (voice)
May Parker (voice)
Martin Li / Mister Negative (voice)
Mary Jane Watson (voice)
Shocker / Additional Voices (voice)
Norman Osborn (voice)
Jefferson Davis (voice)
Miles Morales (voice)
Rio Morales / Additional Voices (voice)
Silver Sable / Additional Voices (voice)
Tombstone (voice)
Taskmaster / Additional Voices (voice)
Dr. Morgan Michaels / Additional Voices (voice)
Electro / Additional Voices (voice)
Rhino / Additional Voices (voice)
Scorpion / Additional Voices (voice)
Vulture (voice)
Harry Osborn / Additional Voices (voice)
Black Cat / Additional Voices (voice)
Short Order Cook (voice)
Screwball / Additional Voices (voice)
Detective Mackey (voice)
Hammerhead (voice)
David Obademi (voice)
Additional Voices (voice)
Additional Voices (voice) (as Issac Singleton)
Additional Voices (voice) (as Jon Bentley)
Additional Voices (voice) (as Kris Zimmerman Salter)