Spears auctioned dozens of personal items on eBay, with proceeds going to The Britney Spears Foundation to benefit the Mississippi Hurricane Recovery Fund.
On Saturday, she posted a message on her Web site saying she was "concerned that some of you might be confusing this bra with something that it's not."
The 23-year-old pop star said she wore the white stone bra — bids had risen to over $47,000 on the auction Web site — during her HBO special but not during her performance of "... Baby One More Time."
"I feel the correct thing to do is remove this item from the auction because I don't want any of you to feel misled," she said.
It was the second time the bra had been removed from the auction. Earlier, an eBay staffer pulled it because it violated the Web site's policy of not selling used undergarments.
The decision was reversed, though, because the bra fit under the category of memorabilia.
The auction otherwise concluded with a "I Have the Golden Ticket" tank top going for $1,200, her ottoman selling for $510, her Chanel high heels for $770, her two-piece sectional sofa for $3,050 and her red-tab jeans for $4,001.
Spears, who is married to her former backup dancer Kevin Federline, gave birth to the couple's first child, a son, last month.