Biden, while campaigning for Hillary Clinton in Toledo, Ohio, appeared to be exhausted by the Republican nominee saying he was "not gonna say anymore" when the crowd cheered and prompted him to continue.
"I'll get myself in trouble and say something like, I'd like to take himself behind the gym if I were in high school," Biden said, referencing his remarks in Pennsylvania Friday.
"I mean, all kidding aside, wouldn't you? Biden asked the audience. "I mean, for real. Can you imagine a guy in the locker room talking that way? And your sister's out there watching the game. Not a joke."
"If I were in high school ... I want to make it clear I understand what assault is. I'm not in high school. If I were in high school," Biden said, finishing his riff punching one hand into the other.
On Friday, Biden at a Clinton rally was slamming Trump's recent comments, calling them "the textbook definition of sexual assault."
He added: "The press always ask me, 'Don't I wish I were debating him?' No, I wish we were in high school -- I could take him behind the gym. That's what I wish."
Biden also challenged Trump's fitness for office, claiming: "One thing he's demonstrated -- he doesn't know anything."
"He says these things, and you're looking -- it's like a sitcom. Like, whoa! Where'd that come from?" Biden said with a smile.
This was the second of three stops on the campaign trail Monday for Biden in the swing state of Ohio.
Earlier, while visiting a Clinton campaign office in Cleveland, Biden warned of a "nightmare" presidency if Trump were elected.
"We all know the nightmare of a Trump presidency," Biden said while surprising a group of volunteers at a campaign office for Clinton. "I've given up on talking about Trump."
Biden, while making clear his disdain for the Republican nominee, was confident in telling the volunteers, "Hillary's going to win this election," and urged them to continue their work in the final days of the campaign.
Biden has one final event in Dayton, Ohio, Monday.