Originally released in 1982, the illustration depicting the title character and Black Widow in a spider's web sold for $101,575 (£70,200).
"The Black Widow has a desperate grasp on Matt Murdock in this hauntingly memorable cover illustration by Frank Miller from his groundbreaking run on the series that thrust him into Marvel superstardom," read Heritage Auctions Galleries' description of the piece.
"At once menacing and intricately beautiful, Miller spins a fascinating design, with the intricate lacing white-line work juxtaposed against stark black shapes."
Frank Miller is renowned for his gritty cover designs, rendered in complex black and white patterns of light and shadow. His notable works include The Dark Knight Returns, Sin City and 300.