Monye's omission is the only change from the squad that defeated Fiji 54-12 at Twickenham at the weekend, with Gloucester's Charlie Sharples retaining his place having scored two tries in the first autumn international of the series.
Harlequins star Monye misses out despite registering a try on his first England appearance in more than two years against Fiji.
"It is tough on Ugo, who misses out this time, but Chris has been excellent in training and Charlie is rewarded for his performance at the weekend," said Lancaster, who names his team tomorrow. "We had to make some tough decisions within the 23 and we are in a good position to be able to select from a largely fit squad."
Prop Alex Corbisiero and centre Jonathan Joseph have failed to prove match fitness and have been sent back to their clubs, along with Monye and 10 other players.
England have defeated Australia in their last two matches and Lancaster will be hoping Ashton repays his faith in the winger, who has scored three tries against them in three Tests.
Meanwhile Australia must respond to their humiliating 33-6 loss in Paris on Saturday without flanker David Pocock, who has been ruled out with a calf strain. The Wallabies will also miss lock Kane Douglas, who featured in the heavy loss to France but has since picked up a knee injury.