"The appeal has been dismissed by the Supreme Court," Tamin Husain Shawan said, adding that the decision, made by the full seven-member Supreme Court bench of judges, was unanimous.
The central bank, which is nominally independent from the government, removed Yunus on March 2 on the grounds that he had failed to seek its approval when he was reappointed indefinitely in 1999.
The High Court upheld this order in a March 8 ruling, with judge Muhammad Mamtaj Uddin Ahmed saying that the central bank's sacking order was legal and that Yunus had also exceeded Grameen Bank's mandatory retirement age of 60.
Backed by a high-profile international lobby group, Yunus, 70, has defied the sacking order by returning to work at Grameen Bank's headquarters and launching a legal battle to keep control of his organisation.
Supporters say he has been victimised by Prime Minister Sheikh Hasina, who he crossed in 2007 when he set up a political party during a period of military rule.
In December, following the release of a Norwegian TV documentary critical of Yunus, Prime Minister Hasina accused him of "sucking blood from the poor" and pulling a financial "trick" to avoid paying tax.
Having exhausted his personal legal options, Yunus's only hope now resides in a separate legal case lodged by nine Grameen Bank board members in the Supreme Court that also seeks his reinstatement.
The case may be heard on Wednesday at the Supreme Court, lawyers said, but few observers expect it to be successful.
Attorney General Mahbubey Alam said Yunus's legal battle was now over.
"The Supreme Court today has upheld the verdict of the High Court," Alam told AFP after the hearing, adding it was "highly unlikely" the verdict on the Grameen Bank board members' appeal would be different.
"After this verdict by the country's highest court, Yunus's position as managing director of Grameen Bank is illegal," he said.
The Supreme Court took into consideration in its ruling the fact that Yunus, at 70, is 10 years past the mandatory retirement age for Grameen Bank officials, Alam said.
"There is no possibility of a compromise," he said, adding that "a person can retire but an institution will not die."
Grameen Bank's government-appointed chairman, Muzammel Huq told AFP that he would call a board meeting after he had received a full copy of the Supreme Court verdict.
According to Grameen Bank's internal rules, the board, nine out of 12 of whom support Yunus's legal battle to stay on at Grameen, has the right to appoint a new managing director.
As well as being sacked, Yunus has been vilified in the Bangladeshi press and summoned to court three times in cases nominally connected to Grameen. The bank has become the target of a government probe.
He will make a statement on the Supreme Court verdict at 5:30 pm (1130 GMT) Tuesday, Grameen Bank spokeswoman Jannat E. Quanine said.
Analysts say Grameen's huge influence in Bangladesh and its move into solar panels, mobile phones and other consumer goods has triggered the government's envy.
His sacking has sparked street protests in Bangladesh and widespread condemnation from overseas, with the US government warning earlier this month that bilateral ties would be affected if an amicable resolution was not found.
Grameen Bank, which is 25 percent state-owned and employs 24,000 people, provides credit to eight million borrowers, the vast majority of them women living in rural villages.
Its work has been copied in developing countries around the world.