The group, which was founded in Washington in 1986, is the fastest growing food chain in the United States and launched its first UK venue in central London last year.
The O2 outlet will be its fifteenth branch on these shores with the company rapidly expanding.
Richard Collier, property director at Five Guys UK, said: "Five Guys' expansion across the UK is highly dependent on the locations chosen, and The O2 was on our list of locations from day one.
"It gives us the buzz and atmosphere of London in a location that provides access to a unique catchment area."
The branch will open near to The O2 cinema on November 9, in place of the Armadillo bar and restaurant.
As well as burgers and fries, Five Guys specialises in hot dogs and grilled sandwiches.
John Eckbert, director of Five Guys UK, said: "We are excited to be opening our next central London venue at The O2, which is home to some of the best eating and drinking locations in London. As an incredible hub for entertainment that draws visitors from all over the UK, we are pleased to offer visitors the perfect pit stop to refuel and discover our vast selection of fresh burgers and signature Five Guys style fries."