The budget carrier’s inaugural flight took off on Tuesday (November 13).
The three-times weekly service will operate on Tuesdays, Thursdays and Saturdays.
Fares lead in from £17.99.
Kharkiv is renowned for its cultural attractions and is a recognised centre for science.
Owain Jones, Wizz Air UK managing director, said described Kharkiv as a “truly undiscovered gem of major cultural significance”.
“This new service is the most recent of seven exciting UK route launches in the past month as Wizz Air continues to expand its UK operations and bring low cost travel opportunities to UK customers.”
Wizz Air now bases nine Airbus A320 and A321 aircraft at Luton.
Lead-in fares are one way, hang luggage only including taxes and non-optional charges.