The Manning brothers will join the broadcast from a remote location for 10 games in each of the next three seasons. Other celebrities, including current and former players, will join the broadcast.
The Monday Night Football Megacast will air on ESPN2, with the potential to air on other Disney properties, including ESPN+.
The Monday Night Football Megacast airs in addition to the network's traditional telecast, which broadcasts on ESPN and/or ABC each week and features Steve Levy, Brian Griese, Louis Riddick, Lisa Salters and John Parry.
"Offering multiple Monday Night Football viewing options for the next three seasons continues our innovation efforts and provides additional value for our fans," ESPN Chairman Jimmy Pitaro said in a statement Monday.
"Peyton and Eli will bring a different approach, delving into conversation about broader, big-picture topics while also honing in on the game, much like fans do when watching with their family and friends."
ESPN said the telecast will be produced in conjunction with Omaha Productions, Peyton Manning's production company he launched in February.
"This partnership with ESPN and The Walt Disney Company reflects an ongoing, shared commitment to offering fans fun, innovative content," Peyton Manning said.
The inaugural Monday Night Football Megacast will air when the Las Vegas Raiders host the Baltimore Ravens on Sept. 13.
Peyton, 45, spent 18 seasons in the NFL. The 14-time Pro Bowl selection, seven-time All-Pro, five-time MVP and two-time Super Bowl champion retired in 2015. He will be enshrined into the Pro Football Hall of Fame on Aug. 8.
Eli, 40, spent his entire 16-year NFL tenure with the New York Giants. The four-time Pro Bowl selection also won two Super Bowls before he retired in 2019.