William Stevenson, 77, was married to Jill Biden from 1970 to 1975.
A state grand jury this month charged Stevenson with killing Linda Stevenson, 64, who was found unresponsive at their home in Wilmington on December 28.
He has remained in custody since he was charged on February 3 with murder, unable to post the $US2 million ($A2.8 million) bail.
Stevenson appeared at Tuesday's court hearing in Wilmington on a remote video camera.
His lawyer, a public defender, did not immediately return messages seeking comment. His next court hearing is set for March 16.
William Stevenson founded the Stone Balloon, a popular music venue in Newark, Delaware, in the early 1970s.
Linda Stevenson ran a bookkeeping business and was described in her obituary as a family-oriented mother and grandmother and a Philadelphia Eagles fan. The obituary does not mention her husband.
"One hug from her and all your worries would disappear," her daughter, Christine Mae, wrote in a Facebook post.
"The pain of losing her is paralyzing and the emptiness in my heart is an abyss."
Jill Biden married US Senator Joe Biden in 1977. He served as US president from January 2021 to January 2025.
Jill Biden's spokesperson has said she has no comment on the Stevenson case.
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