Established captain John Terry will not start the match against the world and European champions and with Rio Ferdinand and Steven Gerrard - two players who have captained the side under Fabio Capello - both absent, Terry's Chelsea team-mate has been handed the honour.
Terry is currently being investigated by the Football Association following allegations that he racially abused QPR defender Anton Ferdinand but will captain the side in Tuesday's friendly against Sweden, also at Wembley.
Capello said he had chosen not to start Terry against Spain before the racism row erupted. When asked why he did not drop Terry entirely, the manager replied: "For me he is innocent until proven guilty. Sorry I can't speak about this at the moment.
"I decided that he will not play this game against Spain because I wanted to see something different with the two centre-backs.
"I decided everything before what happened, and I don't like to speak about this at the moment."
Lampard, who will win his 90th cap when he leads England out on Saturday, said: "It will be my proudest moment in an England shirt. It's a very special moment for me and I'm absolutely delighted."
Meanwhile, Aston Villa forward Gabriel Agbonlahor has been forced to withdraw from the England squad due to a hamstring complaint.
Agbonlahor was included in Capello's squad after scoring five goals and claiming four assists in 11 Premier League games this season.
However, the 25-year-old will be denied the chance to audition for the role of Wayne Rooney's replacement for the opening three games of Euro 2012.
An FA statement read: "Gabriel Agbonlahor has this morning returned to Aston Villa.
"Having been assessed by the England medical staff, Agbonlahor is unable to train today at Wembley and has returned to his club."
Rooney, absent from the squad to face Spain and then Sweden on Tuesday, is suspended for the group stage of Euro 2012, although the FA has appealed against the ban.
With Agbonlahor now absent, Darren Bent, Danny Welbeck, Danny Sturridge and Bobby Zamora are in contention to start against Spain at Wembley.
Phil Jagielka missed training on Thursday with a fractured toe but the Everton defender emerged at Wembley on Friday to join his team-mates.
"He played normally today (Friday)," said Capello. "I spoke with him. I asked him if he is ready to play. He told me yes."