England defender John Terry has been ruled out of Tuesday's World Cup qualifier against Ukraine due to an ankle problem.
Terry suffered the injury during Friday night's 5-0 win in Moldova and England finished the game with ten men after the Chelsea defender limped off in the closing stages.
He received treatment on Saturday and England remained hopeful he could take his place in the heart of the defence at Wembley, but the centre-back has lost his fitness race.
The Football Association confirmed Terry's withdrawal in a statement. It read: "Following further treatment from the England medical staff today, John Terry will not be available for the World Cup qualifier against Ukraine on Tuesday.
"Terry suffered an ankle injury in the game with Moldova and has returned to his club.''
Chelsea team-mate Gary Cahill is likely to step up to make his first appearance at Wembley since he suffered the broken jaw against Belgium which ruled him out of Euro 2012, and replace Terry.
The other player in contention to partner Joleon Lescott is his former colleague at Everton, Phil Jagielka.
Left-back Ashley Cole had already been ruled out with the ankle injury which prevented him from playing in Chisinau. But Leighton Baines proved a capable deputy against Moldova and scored England's final goal.
Frank Lampard believes England possess the strength in depth to fill the gap created by the injury to Terry.
He said: "It is a blow for John personally and for the team because he is an inspirational player and a fantastic centre-half. He is a top player and experienced in these qualifying games. We saw his ability through the Euros in the summer and the other night and a big player, you always miss.
"But it's squad football and we are fortunate enough to be blessed with fantastic back-up, good centre-halves like Cahill and Jagielka. Whoever comes in are experienced top players. Whenever you lose a big player, you need the squad to step up and do the business and I'm sure we've got that.''