The film, starring Daniel Craig as gentleman detective Benoit Blanc, will have a gala screening at the Royal Festival Hall at the Southbank Centre on Sunday October 16.
Former Bond star Craig will attend alongside director and producer Rian Johnson.
Also in attendance will be Edward Norton, Janelle Monae, Leslie Odom Jr, Kate Hudson and Madelyn Cline, all new to the franchise, with more names to be added.
Simultaneous preview screenings will also take place at additional cinemas across the UK.
Glass Onion: A Knives Out Mystery will see Blanc travel to Greece to peel back the layers (arf arf) of a case involving a new cast of colourful suspects.
Johnson said: ‘I’m thrilled to be back at LFF with Glass Onion, and it’s an honour to be closing the festival.
‘A proper whodunnit really does belong in London, so it feels a bit like coming home!’
Tricia Tuttle, director of the LFF, said: ‘Rian Johnson’s Knives Out was a major hit when we hosted the European Premiere at the Festival in 2019.
‘Our audiences adored the film’s wit and that it tipped its very stylish hat to the wonderful tradition of British onscreen sleuths.
‘And here, Rian Johnson strikes gold again with the help of the year’s hottest ensemble cast. Like its predecessor, Glass Onion is entertaining and culturally literate in equal measure, making some hilarious, razor sharp observations about the world we live in.
‘The European Premiere of the Glass Onion: A Knives Out Mystery will bring the house down as the Closing Gala of the 66th BFI London Film Festival!’
Knives Out, which co-starred Chris Evans, Ana De Armas and Christopher Plummer, also had its European premiere at the LFF in 2019, with Johnson and the cast walking the red carpet.
Netflix won the distribution rights to the budding franchise for a chunky $469m (£330m), under the condition that two sequels – both starring Craig – be produced.
The 66th BFI London Film Festival takes place from Wednesday October 5 to Sunday October 16.
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