The campaign, created by Euro RSCG London, breaks in cinemas on 26 March.
The 40-second spot shows the fairy flying around London's attractions.
She picks up a food wrapper beside a couple too engrossed in each other to notice her, then she gets caught up in a plastic bag, which lights up like a lantern as she tries to escape.
She also struggles to remove chewing gum from the Millennium Bridge and picks a bottle out of a fountain in Trafalgar Square.
The voiceover, by John Hurt, questions whether viewers believe that fairies come down at night to clear the streets. The ad closes with the fairy flying away as dawn breaks over London.
The creative aims to instil the message that cleaning up London is up to its inhabitants. The ad ends with the question: "Does a helpful litter sprite clear away your mess at night?"
Ken Livingstone, the Mayor of London, said: "Rubbish left on our streets and other public places is spoiling the beauty of our capital city and initiatives like this advertising campaign are essential if London is to remain a great city to live and work in."
The ad was written and art directed by Olly Caporn and directed by Seb Edwards through The Brave Film Company.
The cinema ad will be backed by 40-second radio ads and six-sheet posters in a media strategy devised by Pawson Media.