- Iron Man - Tom Kenny
- Captain America - Roger Craig Smith
- Thor - Travis Willingham
- Hulk - Troy Baker
- Spider-Man - Yuri Lowenthal
- Gwen Stacy - Erica Lindbeck
- Miles Morales - Nadji Jeter
- Ghost Rider - Kirk Thornton
- Ant Man (Hank Pym) - Patton Oswalt
- Wasp (Janet Van Dyne) - Krystina Alabado
- Ant Man (Scott Lang) - Grant George
- Black Widow - Vanessa Marshall
- Hawkeye - Alan Tudyk
- Nick Fury - Dave Fennoy
- Wolverine - Steve Blum
- Cyclops - Quinton Flynn
- Professor X - Simon Callow
- Laura - S
- Harry Osborn - Josh Keaton
- MJ - Laura Bailey
- Aunt May - S
- J.J. Jameson - J.K. Simmons
- Black Cat - Jennifer Hale
- Otto Octavius - Clancy Brown
- Norman Osborn - Roger L. Jackson
- Curt Connors - Jason Isaacs
- Adrian Toomes - Richard Horvitz 
- Morrie Bench (Hydro) - Norman Reedus
- Human Torch - Matthew Mercer
- Blade - Keith David