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- The film shows a bond beginning to form between Scarlet Witch and Vision, who were actually married for a time in the comics.
- Chris Evans injured his arm muscles while filming the iconic shot of Captain America holding back a helicopter with his hands. Evans said, "That shot is a little bit of bicep porn. 'Zoom in on the biceps' - that's what the script said. [?] Kevin Feige didn't airbrush my biceps, that's me. It's not a utilitarian shot, it's a shot where you're trying to look heroic. That position (of holding the helicopter with one hand and the ledge with another) is actually a very unnatural position to use to stop something but we used it because I have to flex my bicep, you are trying to make the scene look great. And I genuinely messed my arm doing that shot because of the strain." Robert Downey Jr. joked that the film-makers didn't mind that Evans hurt himself because of how great the shot turned out and they have been using it to promote the film ever since. Anthony Russo revealed that the iconic shot almost did not happen because on the day of the film-making there was a miscommunication with the costume department and Evans came in wearing a thick jacket. They immediately sent him back to change because then they wouldn't have been able to film the shot of his muscles bulging as intended in the script.
- Chris Evans revealed that he was hesitant to take the role of Captain America early on, but now considers it one of the best decisions he has ever made.
- Tom Holland is the youngest person to portray Spider-Man/Peter Parker on screen at 19 years old.
- Robert Downey Jr. and Chris Evans described the dynamic between Iron Man/Tony Stark and Captain America/Steve Rogers as a marriage, saying, "We love each other, but it's explosive. You're working toward the same goal, but you have very different approaches to it. It's blurry, and that's what makes it great; no one's right, no one's wrong. It's going to make it even harder for them to come to an agreement."
- When the production of the film wrapped up, Sebastian Stan and Anthony Mackie raced each other while in their super hero suits.
- Tony Stark (Robert Downey Jr.) remarks how attractive Aunt May (Marisa Tomei) is. Downey and Tomei actually had a relationship in the 1990s, and appeared in two other films together: Chaplin and Only You.
- The day before filming a fight scene with Robert Downey Jr., Sebastian Stan sent him a video of himself doing intense bicep curls in front of the decapitated head of an Iron Man suit. He attached the message, 'Looking forward to our scene tomorrow Robert.'
- Tom Holland commented on how intimidated he was when he came to screen test for Spider-Man, saying that Robert Downey Jr. took him aside and said, "Listen, I remember my screen test for Iron Man. I remember how terrified I was. Just think of it as an audition. It's nothing too scary. If you get it wrong, we'll just start again. No pressure."
- William Hurt previously appeared in The Incredible Hulk as the on-screen father of Liv Tyler's character. That makes this film the second time that Martin Freeman has worked with Tyler's on-screen father. Previously, Hugo Weaving played her father in the [Lord of the Rings] films, and returned to play the role again in the [Hobbit] films. Weaving and Richard Armitage had previously appeared together in Captain America: The First Avenger.
- T'Challa had a much smaller role in the earlier drafts and did not even appear in costume, as the writers intended to put more focus on Spider-Man and save Black Panther's origin for his own movie. However, when it looked like Marvel would not be getting permission to use Spider-Man, Black Panther's role was beefed up significantly. By the time they did get to use Spider-Man, T'Challa had already become so integral to the plot that they decided to leave his role as it is and give Spider-Man a smaller part.
- Jim Rash has a cameo near the beginning of the film. Rash worked with the Russo Brothers in [Community]. Another [Community] star, Danny Pudi, made a cameo appearance in Captain America: The Winter Soldier.
- [?] Chad L. Coleman was considered for the role of Black Panther.
- Logan Lerman and Dylan O'Brien were considered for the role of Peter Parker. The reported six finalists were Tom Holland, Asa Butterfield, [?] Judah Lewis, [?] Matt Lintz, [?] Charlie Plummer, and [?] Charlie Rowe before Holland was cast. The finalists for the role did separate screen tests with Chris Evans and Robert Downey Jr., with the intent of seeing whose chemistry fit the best with both Evans and Downey.
- Robert Downey Jr.'s personal trainer Eric Oram stated that the trick to pitting Captain America against Iron Man "is to show Iron Man using the minimum force necessary to win the fight, and not to look like he's trying to go wild and kill somebody."
- Alfre Woodard has a cameo as Miriam Sharpe, the mother of an American citizen killed in the battle of Sokovia. Woodard was suggested for the role by Robert Downey Jr., before Marvel Studios learned of her casting as Mariah Dillard in [Luke Cage].