However, officers said they would "remain committed to bringing the perpetrators of this crime to justice".
It follows a robbery at Lloyds TSB Scotland in Dunearn Drive, Kirkcaldy at about 10:40 on Friday 29 January 2016.
Two men entered the bank and threatened staff with a handgun before leaving with a four-figure sum of money. Police are appealing for witnesses.
Detectives previously released CCTV images and footage of two men they want to trace in connection with the robbery.
The first man is white, in his late 20s or early 30s, with dark stubble on his face. He was wearing a dark-coloured baseball cap and dark-coloured clothing.
The second man is white, 5ft 9in, about 30 years old, of medium build and with stubble.
He wore a dark-coloured baseball cap, a dark blue/grey hooded top and dark-coloured trainers.
He was also wearing dark blue jogging bottoms with a distinctive white-coloured motif that read MORDEX and partially ran down one leg. Mordex is a Polish brand typically associated with bodybuilding.
The first man was seen riding a mountain bike and the second a grey and red mountain bike on Alford Avenue before the incident, and in the area of Cawdor Crescent a short time after the incident.
Both were also seen carrying green carrier bags.
Det Insp Graeme Dursley, of Police Scotland, said: "One year on from the robbery, we've exhausted almost all possible lines of inquiry but we remain committed to bringing the perpetrators of this crime to justice.
"We've conducted significant investigations in the area including speaking to over 400 local residents, reviewing CCTV and collecting forensic evidence at the scene.
"The support from the local community for both our investigation and the bank staff following this terrifying incident has been tremendous and I want to thank everyone who came forward.
"We've issued numerous appeals to the public, including releasing CCTV footage and an appeal on BBC's Crimewatch Roadshow, but I believe that someone in the community may still hold vital information.
"Anyone with information that may be relevant, who saw the suspects in the area and has not yet spoken to us, or who recognises their descriptions is urged to get in touch as soon as possible."
Police have 'exhausted almost all possible lines of inquiry' one year after an armed robbery in Fife.