The groups set out from Hornsea and Southport yesterday, carrying batons containing a map of the route.
The Hornsea cyclists headed for Hull, where the baton was due to be signed by the city's Lord Mayor, Coun David Gemmell.
A total of six teams of baton-carrying cyclists are taking part in the celebrations and are due to meet at Barnsley football ground, Oakwell, on Sunday, where a plaque will be unveiled to commemorate trail founder Robin Norbury, who died this year.
Coun Gemmell said: "The Trans Pennine trail is a fantastic route to explore the North of England, passing through some of our prettiest countryside and historic towns and cities.
"It has been developed so that the terrain is suitable for everyone."
The concept of a trail linking the Irish and North Seas began in 1987 and an inaugural cycle ride and walk took place between Southport and York in 1989.
Sections of the route opened gradually until the trail was finally completed in September 2001.