- J. Jonah Jameson has high blood pressure and medication to take, whilst Miss Brant has a charge from his wife to make sure he doesn't lose his temper, a long history of being treated like his dogsbody and an intercom with a loud buzzer. Revenge is had.
- A buzzer so loud, that when the desk gets rattled every time, it's hard to tell if it was because of the vibrations or Jameson's startled jumps.
- While probably unintentional, MJ's line to Peter accusing him of a possible relationship between him and Gwen Stacy. "How come you never mentioned her? She's your lab partner, you saved her life, she thinks you're a genius and she had her polished fingernails all over you?" Just the way she finishes with the "polished fingernails" bit makes it sound like Arson, Murder, and Jaywalking, as if Gwen having polished fingernails is part of what's wrong. Then again, MJ is sort of the tomboy to Gwen's girly girl.
- The scene where Peter Parker, under the possession of the symbiote, conclusively disproves both that Evil Is Cool and Evil Is Sexy by strutting down the street like someone out of Saturday Night Fever with a spectacularly daft Emo haircut, convinced he's the hot-stuff despite the looks of confusion, bemusement and disgust from almost every woman he crosses paths with.
- I distinctly remember a couple of women looking intrigued. One was even staring at his ass. When I realized this, I could not stop laughing for 3 minutes straight.
- Dr. Connors watching the sample of symbiote biomass bumping Peter's blood cells to the beat of the background music.
- And then there's the dance scene in the jazz club, where he starts off playing piano (out of nowhere, really) and then says "Double time!", jumps over the piano, walks across the bar (stealing a guy's drink in the process) before doing a ridiculous and impossible slide across the room on a chair. After taking off his jacket (the force of which blows back Gwen's hair... somehow) and throwing it at the stage, he does a stupid little to-and-from slide before saying, in a completely serious voice, "Now dig on this..." before air-thrusting and snapping his fingers to the tune of the music. It's so ridiculously stupid, it's funny.
- During the final battle, Peter and Harry are flying downward on his glider trying to catch a falling Mary Jane. Peter asks if it can go faster, Harry responds with 'Hang on!'. Peter aptly notes 'To what!?'
- Also when Harry says 'Hey Pete! Am I interrupting!?' when intruding on a Peter/MJ moment.
- While watching the final battle, JJJ gets conned by a kid. See it here.
- "How's the pie?" "...SO GOOD."
- Most of Eddie Brock's moments before he starts to have his Sanity Slippage.
- Jameson walks in on Peter trying to flirt with Betty Brant on his desk.
Jameson: Miss Brant, that's not the position I hired you for!
- From the DVD commentary when Flint Marko's mugshot is revealed to Peter and Aunt May:
Thomas Hayden Church: It was that guy. He was also in George of the Jungle.
- When Spider-Man confronts Sandman in the armored truck, he initially thinks he's just dealing with a standard criminal. He realizes differently when he takes a swing and his fist gets stuck in Sandman's chest.
- After the fight, Spider-Man cleans some of the sand out of his costume and asks, "Where do all these guys come from?"