The New York Republican is only the sixth lawmaker in history to be removed from the lower chamber of Congress, and the first since 2002.
His tenure was marked by multiple lies about his past and allegations of fraud – all revealed after his election.
Friday’s was the third attempt to oust Mr Santos after two previous votes failed.
The 35-year-old from Queens made a quick exit from the Capitol before the vote ended as its outcome became clear, rushing past a swarm of reporters and into a waiting SUV.
“As unofficially already no longer a member of Congress, I no longer have to answer a single question from you guys,” he said.
Lawmakers backed the expulsion resolution 311 to 114, with 206 Democrats and 105 Republicans voting in favour.
Scattered applause was heard across the House chamber after the measure was adopted.
Over 11 months in office, Mr Santos faced an endless stream of controversy and countless calls to resign from members of both parties.
His troubles began shortly after winning election to the House in November 2022, when the New York Times reported he had lied about a Wall Street career, his college degrees and having Jewish ancestry.
Since then, the allegations have only piled up. He has been accused of a range of fabrications, from scamming Amish dog breeders in Pennsylvania to claiming his mother died in the 9/11 terror attacks.
In May, he was charged with 23 felonies, including wire fraud, money laundering and theft of public funds. He denies the allegations and is awaiting trial.
But the final blow came last month, when the House ethics committee found he had exploited “every aspect of his House candidacy for his own personal financial profit”.
Among its many allegations, the panel accused him of spending campaign money on Botox treatments, credit card debt, OnlyFans – a platform where users pay for content, including pornography – and trips to the Hamptons seaside enclave in New York.
Expulsion votes are rare in Congress and require the backing of two-thirds of the House.
Two previous efforts to remove Mr Santos failed, after some lawmakers argued it would set a bad precedent to remove someone who had not been convicted of crimes or tried in a court of law.
Jim Jordan, a Republican who voted against the expulsion, told the BBC he worried “who’s next”.
“The voters elected him,” he said. “You’ve got to be careful in taking a vote to kick out of Congress someone the voters sent to Congress.”
A group of four New York Republicans, all elected alongside Mr Santos, had been trying to get him ousted.
“The precedent that is set is that we hold members of Congress to a higher standard,” said one, Anthony D’Esposito.
“It shouldn’t have come to this,” he added. “He should have held himself accountable. He should have resigned.”
In the days after the release of the ethics committee’s report, Mr Santos had refused to quit, slamming colleagues online and daring them to remove him.
“This place is run on hypocrisy,” he told reporters earlier this week. “If they want me to leave Congress, they’re going to have to take that tough vote.”
Constituents in his district welcomed the news, with one saying only “good riddance” when asked for their reaction.
Jody Kassfinkel, who campaigned to have Mr Santos removed, told the BBC his expulsion was “a win for democracy”.
“We knew this was the only way to go because this man has no shame and he was not going to resign on his own,” she said.
What happens next?
As soon as the vote was gavelled out, Mr Santos officially became a former member of Congress.
His official website was taken down, his staff phones now go to a generic voicemail and the nameplate outside his office – where some people stopped by to take selfies on Friday – was removed.
A sign attached to the doorway says “Yes! We’re open” – but there were no signs of life inside except for a staffer who briefly exited to pick up flowers left at the entrance.
The New Yorker no longer has the ability to vote on legislation or to rely on his government health benefits, and is not eligible for a congressional legislative pension.
He can still dine, however, in the exclusive House restaurant, exercise in the Capitol gym or borrow books from the Library of Congress – all privileges afforded to former members of Congress.
New York Governor Kathy Hochul has 10 days to call for an election, which will likely take place next February.
With the Republican House majority shaved to just eight seats, Democrats have a good shot of filling the vacancy.
It caps a stunning downfall of a man who scored an upset victory in a Democratic-leaning district and became the first openly gay Republican elected to Congress.
With a federal fraud trial looming next year, some have speculated he will sign a plea deal with prosecutors to avoid prison time, as he did with a case in his native Brazil earlier this year.
If not, he faces a maximum penalty of at least 20 years behind bars.
Readers’ comment on why Africans bad politicians are not call to account.
Additional reporting by Pratiksha Ghildial and Nadine Yousif.
Credit BBC