The last scheduled flight will leave the City Centre Airport on June 29.
Northern Air, which flies to Peace River, is moving to the Edmonton International Airport after its licence expires at the end of the month.
"We're hopeful that our customer base will still follow us out there," said Rob King president of Northern Air, the only scheduled carrier left at the airport.
King supported the long-standing battle to save the airport, but said the fight is over.
"For those that are fighting that, it's an uphill battle," he said. "But nonetheless, they have fought and I don't know how successful they'll be. I wish them luck."
More than a half-dozen businesses will make the same move later this year, said Traci Bednard, spokesperson for Edmonton International Airport.
"We have the services and we do have the space available," she said. "These are still our tenants, they're business partners to us. So, we want to support their success."
Canadian Helicopters and STARS air ambulance will move later this year.
Still, other firms including charter airlines, airplane repair shops and hangars say they won't move until the downtown airport closes for good, which could be as early as 2014.