Assassin's Creed: Brotherhood is the third game in the series and was released on November 16 (USA) and November 19 (UK), 2010. The game is a continuation of Ezio Auditore's story, the Assassin's Creed II protagonist, as he returns to the city of Monteriggioni. The day before his sister's surprise birthday party for him he spends the night with Caterina Sforza, waking up to find the city besieged by Cesare Borgia.



Ezio joins the attempt to fight off Cesare's troops but during the raid his uncle Mario is killed by Cesare, who then steals the Apple of Eden, leaving Ezio to escape the city without him. Wanting to exact justice and retrieve the Apple, Ezio travels to Rome in order to liberate its people from the influence of the Borgia family. To do so, Ezio recruits new assassins, as well as forming alliances between the various factions of the Roman underground. There are several times that Ezio goes to the Castel Sant'Angelo to assassinate Cesare and Rodrigo Borgia but they escape.



When Ezio comes back to the Castle, he sees Cesare kill Rodrigo by forcing him to eat a poisoned apple, which Rodrigo himself had poisoned in an attempt to kill Cesare. Ezio chases Cesare to the Apple of Eden and manages to claim it. Cesare is arrested but he says that no man can kill him and chains will not hold him for long. Cesare is placed in a high security prison in Viana, Spain, however he manages to escape with the help of his followers, including Micheletto. Micheletto is eventually killed by Cesare after a bar room fight that leads to where the memory starts in the game. Ezio kills Cesare, throwing him off the castle walls; 'leaving him in the hands of fate'. After that, Desmond, Lucy, Rebecca and Shaun find that the Apple of Eden was hidden under the Colosseum.



They search for the Apple and finally find it, then Juno, another of "Those Who Came Before," controls Desmond with the Apple and makes him stab Lucy. He then falls into a coma while Lucy's condition is unknown.



For the first time in the series, the game features online multiplayer. Players play as Abstergo employees, who, through the use of the Animus, take on the genetic memories of Renaissance Templars in various game modes. The Da Vinci Disappearance DLC ends inside one of the 5 temples and gives GPS coordinates that coincide with a lake in New York. It is hinted that the Apple of Eden has already been hidden in a temple by Ezio and the DLC takes place 3 years before the end of Brotherhood. After the final memory, Desmond is still in a coma and hears unfamiliar voices of the new assassins he has yet to meet. They say they have found what they need and hint at dispatching an assassin team to the new found temple.