This will be Klitschko's 17th world title fight, but his first for nine months since he stopped Ruslan Chagaev in the ninth round in Gelsenkirchen in June 2009.
The 33-year-old Ukrainian is 15 kilogrammes heavier and 15 centimetres taller than his American opponent who has suffered just one defeat in his career, but Klitschko is convinced of victory at Düsseldorf's Espirit Arena.
"He wants to take away my titles and I will not permit this," said Klitschko.
"He is dangerous and fast ... He is very agile, very athletic and is mentally prepared to take the title away from me," said Klitschko. "Of course I won't allow that, I will be very alert and punchy on Saturday night."
Klitschko has won 53 of his 56 fights with 47 knock-outs, and the last of his three defeats was against Lamon Brewster back in 2004.
In contrast, Chambers beat Alexander Dimitrenko in July 2009 to become the WBO's number one contender and the sole defeat of his career was against Alexander Povetkin in Berlin in January 2008.
Chambers is nicknamed "Fast Eddie," and he hopes to counter Klitschko's size, left jab and his notorious hard-hitting right with hand speed and body movement.
This is the biggest fight of the 27-year-old's career, having won 35 of his 36 fights with 18 knock-outs, and he says he is ready to put Klitschko down and walk away with his titles.
"It's going to be very difficult, but I'm prepared for it," said Chambers, who was hired to spar with Klitschko in 2006.
Some 46,000 of the 51,000 available tickets have been sold for the fight at the football stadium, home of second-division side Fortuna Düsseldorf.