The ad shows Trump -- the Republicans' 2024 standard-bearer -- repeatedly mocking the former UN ambassador and telling a reporter he can win without pursuing her supporters.
"She's gone crazy. She's a very angry person," Trump is heard saying during the 30-second commercial, punctuated with social media posts in which the Republican claims he "will not accept" her supporters and threatens to kick them out of his political movement.
Biden posted the video on social media alongside the caption: "Nikki Haley voters, Donald Trump doesn't want your vote. I want to be clear: There is a place for you in my campaign."
Biden has built up a campaign war chest in excess of $70 million ahead of November's rematch with Trump, more than doubling his predecessor's fundraising haul and giving allies more latitude to spend big on TV advertising.
The president raised an estimated $26 million on Thursday at a New York event with former presidents Barack Obama and Bill Clinton.
Trump has had to divide his time between campaign stops and court appearances as he defends himself against 88 felony counts.
He has also had to dedicate tens of millions of dollars in donations that could have boosted his campaign on legal fees.
But he benefits greatly from free coverage as he turns his many appearances at courthouses into campaign events.
The 77-year-old, who has been indicted four times -- mostly over alleged attempts to cheat in elections -- is expected to raise $33 million from a fundraiser on April 6 in Florida, according to the Financial Times.
Trump secured the Republican nomination at the beginning of March after defeating Haley by wide margins in all the early voting and "Super Tuesday" states -- including her home turf of South Carolina.
But large swaths of Republican primary voters -- upwards of 40 percent in some states -- did not support Trump, raising fears that he will not be able to win the moderates he will need to take back the White House.
The TV spot will run for three weeks in eight key battleground states, targeting Haley voters in predominantly suburban districts where she performed best against Trump.
The ad will also appear across a wide array of digital platforms including those owned by Meta -- parent company of Facebook and Instagram -- and YouTube.
"As Trump loses ground, derides women, and threatens to ban abortion nationwide -- Joe Biden is reaching across the aisle to Haley voters, inviting them to join his growing and winning coalition of voters who will decide this election," the president's campaign said.
