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No I'm afraid you'll have to start again.

 

"Unfortunately for us who have already beaten the game once and back again, the trophies will not be retroactive.

IGN: Will any of them be able to be awarded retroactively?

Arne Meyer: That would have been nice, but it wasn't something we would be able to do. We originally left our save games unlocked and portable so gamers could move them off the hard drive and their existing game elsewhere or show off some of the unlocked content at a friend's house. This type of portability would have allowed gamers to get Trophies unfairly and we couldn't let that happen. To receive Trophies in Uncharted you will have to start a brand new save game without any medals or rewards unlocked."