The 78-year-old performer will no longer appear on stage as the rampant purple-haired housewife, who has become iconic for her boisterous greeting of "Hello Possums!".
Dame Edna will bow out with a special farewell tour, the details of which Humphries has not yet disclosed.
Humphries told Australia's Sunday Telegraph: "Edna will crop up on television I guess, but not in a live show. The fact of the matter is that I'm beginning to feel a bit senior."
Of the character's physical demands, 'It's the best aerobics you could do, leaping around on stage, but it's gruelling when there are other things to do."
Her commercial peak came in the late 1980s when she hosted The Dame Edna Experience on London Weekend Television.
Humphries also revealed that he is also retiring the boorish Sir Les Patterson, his second most popular creation.
Explaining his decision, the legend recalled his parents taking him to see stand-ups who had "outlived their shelf life".
"It was commented that, 'You should have seen him when he was funny'. I want to avoid that being said about me and know that I can't keep doing it," he added.
Watch Dame Edna Everage on This Morning below: