Slipper, who made his debut off the bench against England in Perth in 2010, is one of only three Australians, with George Gregan and Adam Ashley-Cooper, to play at four World Cups.
He has made more World Cup match appearances (21) than any other Wallaby, but the lure of a home tournament in 2027 was a bridge too far.
“It feels like the right time for me to step away from test rugby,” he said.
“There’s a number of good young props coming through who will need time in the saddle leading into what’s going to be an unbelievable experience of a home Rugby World Cup in 2027.”
Wallabies coach Joe Schmidt said Slipper was first and foremost “a great person”.
“He’s incredibly popular in the group and is the ultimate team man.
“I know how much representing the Wallabies means to him and his record speaks for itself with the number of milestones he has achieved at the highest level of the game.”
Slipper’s 151st and last test will be at Perth Stadium on Saturday, with Australia still in with a slim chance of winning their first Rugby Championship in a decade.
But they must beat New Zealand with a bonus point and hope Argentina soundly defeat South Africa at Twickenham in London.