MADRID — The European Parliament has stripped the immunity of Carles Puigdemont, the former separatist leader of Catalonia, clearing the way for Spain to make a fresh attempt to extradite him from Belgium and try him on sedition charges.

The European Parliament said on Tuesday that a majority of its members had voted a day earlier in a secret ballot to remove the immunity of Mr. Puigdemont and two other Catalan members of the assembly who face charges in Spain related to a botched attempt to declare Catalonia’s independence in 2017. Spain’s judiciary has charged that their bid was unconstitutional.

The vote on Monday ended a lengthy battle by Mr. Puigdemont and his colleagues to use their protection as elected members of the European assembly to shield them from prosecution in Spain. Now it is up to the Belgian judiciary to rule on whether Mr. Puigdemont should be sent back to the Spanish capital, Madrid, to stand trial.

“It is a sad day for the European Parliament,” Mr. Puigdemont said. “We have lost our immunity, but the European Parliament has lost more than that and as a result, European democracy too,” he said, adding that this was “a clear case of political prosecution.”

The Spanish government welcomed the vote.

“The problems of Catalonia will not be solved in Europe or by Europe. They have to be solved in Spain by bringing all Catalan forces around the table,” said the foreign minister, Arancha González Laya. The vote showed that the European Parliament had “respect for the work of the judiciary in our country,” she added.

The European Parliament’s decision comes only weeks after regional elections in Catalonia that increased the majority of pro-independence parties in the regional Parliament. Separatist politicians have held control since 2015, but the secessionist conflict has split Catalan society while also remaining a highly contentious issue in national politics.

Mr. Puigdemont and some of his colleagues have been in Brussels since October 2017, shortly after the Spanish central government ousted his regional government for holding a referendum that Spanish courts had ruled illegal and then declaring Catalonia’s independence.

During the past three years, Mr. Puigdemont has successfully fought off attempts to extradite him both from Belgium and Germany, where he was briefly detained during a trip.

In January, judges in Belgium also rejected a request to extradite another former member of Catalonia’s regional government, Lluis Puig, who is facing similar charges in Spain. The Belgian court argued that the Spanish Supreme Court did not have the legal authority to issue an arrest warrant against Mr. Puig, adding that he should be tried in a regional court.

Part of Mr. Puigdemont’s former government, however, stayed in Spain and stood trial before the country’s Supreme Court. Nine Catalans received prison sentences after they were convicted of crimes including sedition and misuse of public funds.

One former Catalan leader, Oriol Junqueras, was also barred by Spain’s highest court from taking his seat in the European Parliament. Both he and Mr. Puigdemont were elected to the assembly in 2019.

The European Parliament’s vote will allow a Spanish judge to reactivate a European arrest warrant against Mr. Puigdemont that was suspended in early 2020, when Mr. Puigdemont and his colleagues took their seats in the European assembly.

The Catalan leaders are not the first members of the European Parliament to be stripped of immunity.

In 2019, the European Parliament stripped the immunity of Jean-Marie Le Pen, the founder of the far-right National Front party in France. It is still reviewing the case of Ioannis Lagos, who was sentenced in Greece last year for his activities with the far-right Golden Dawn party. The Greek government considers Golden Dawn a criminal organization.

The Catalan case has divided politicians in Brussels, many of them loathe to set a precedent of lawmakers being tried over political activity. The removal of Mr. Puigdemont’s immunity was approved by three-fifths of the members of the European Parliament.

It could take months for Belgian courts to rule on Spain’s latest attempt to extradite Mr. Puigdemont and the two other Catalan leaders, Antoni Comín and Clara Ponsatí.

The Brussels Public Prosecutor’s Office is examining the possibility of renewing legal proceedings in Belgium, a spokeswoman for the office said.

Should the Belgian courts block the extradition request, the Catalans would continue to sit in the European Parliament, but without special immunity rights.