Under the watchful eye of Father Silvano Bentivoglio and Dr. Vittoria Vetra (Ayelet Zurer), the CERN scientists start the Large Hadron Collider and manage to capture three vials of antimatter. Immediately afterward, someone kills Father Silvano, and uses his retina to break into the containment room to steal one vial of anti-matter.
In Rome, the Vatican mourns the passing of the Pope. The Vatican staff prepares for the Conclave of the College of Cardinals, which will select the next Pope. Until the Conclave selects a new Pope, the Camerlengo (Ewan McGregor), a Vatican priest and former helicopter pilot, assumes day-to-day control of the Vatican. Reporters, nuns, priests and other faithful all crowd into St. Peter's Square, waiting for the white smoke from the Conclave. But the Illuminati, a 400-year old, underground secret society, kidnaps the four most likely candidates (preferiti) before the Conclave goes into seclusion. The Illuminati threatens to kill them at 8, 9, 10 and 11 PM, and then destroy the Vatican in a burst of light at midnight. A video feed shows the missing anti-matter vial, which will destroy the Vatican and parts of nearby Rome when the magnetic containment field fails after the vial's battery dies.
The Vatican summons Drs. Robert Langdon (Tom Hanks) and Vittoria Vetra from CERN to help them solve the Illuminati's threat, save the four preferiti, and replace the vial's batteries. Langdon listens to the Illuminati message and deduces that the four cardinals will die at the four altars of the Path of Illumination. However, no one knows where these altars are located. Vetra demands Commander Richter (Stellan Skarsgård), the captain of the Vatican's Swiss Guard, to bring Father Silvano's diaries from Switzerland. She hopes the father wrote down the name of whomever he discussed the CERN experiment with and who must possibly be the mastermind behind Silvano's murder. Langdon also demands access to the Vatican Library (something he has requested 10 times already) to see the original copy of a Galileo's banned book which may contain the location of several churches used by the Illuminati as gathering points and which were also the altars. Using the clues from this book, Langdon, Vetra, Inspector Ernesto Olivetti (Pierfrancesco Favino) and Lieutenant Valenti (Victor Alfieri) of the Vatican police race to the first church, only to find the dead body of the first Cardinal underground, branded with the word "Earth".
They find the direction of the church with the second Illuminati altar, but arrive in time to see the next Cardinal die after being stabbed at his chest, branded with the word "Air".
While Vetra studies father Silvano's diaries Langdon and the Vatican officers locate the third church with the third Illuminati altar and try to save the third Cardinal from burning to death, but the assassin appears and shoots and kills all the Vatican officers escorting the professor (including Valenti) and after a brief fight cuts Olivetti's throat, effectively stopping Langdon from saving the Cardinal branded with the "Fire" word.
After escaping from the assasin professor Langdon convinces two members of the Rome police to race with him to the last church of the "Water" altar,
but the assasin murders the two policemen before throwing the fourth cardinal into a fountain. However, Langdon manages to save the last preferiti Cardinal this time, who manages to tell Langdon the location of the murderer's hideout. When Lagdon and Vetra arrive they are confronted by the assasin who spares their lives since "he wasn't paid to kill them" and reveals that his contractors were people from the church. The assasin escapes afterwards and upon entering his car and picking up his payment is killed as the vehicle explodes. Langdon and Vetra find clues to the final victim, who turns out to be the Camerlengo. After arriving at the Vatican using a secret passage, they and the members of the Swiss Guard enter the Camerlengo's room and find him in the floor branded with the vatican church symbol on his chest and apparently about to be shot by Commander Richter of the Swiss Guard.
The Swiss Guard officers shoot at their commander to save the Camerlengo. During the confussion the dying Richter gives Langdon a key to his office. Then the Camerlengo, Langdon, Vetra and the Swiss Guard officers discover the location of the stolen antimatter vial. When they find it, they discover that the battery is about to expire, which would cause the vial to explode and destroy a large part of the city. The Camerlengo takes the vial and uses a helicopter to fly away from the Vatican. Then the priest sets the autopilot and escapes from the helicopter, using a parachute to land. After several seconds the bomb explodes and the Camerlengo lands, being considered a hero by the people present at the Vatican and even as the best candidate to be the new Pope by the members of the Conclave. Meanwhile professor Langdon and Vetra use the key from Richter and watch a video tape that he kept in his office which showed that the mastermind behind the murders of the original Pope, the preferiti and the stealing of the antimatter vial was in fact the Camerlengo and not the Illuminatti. Richter discovered this and while he tried to arrest the Camerlengo the priest branded himself with the church seal and accused the commander of the Swiss Guard of being a member of the Illuminatti and trying to kill him. Langdon shows the video to the members of the Conclave and after the Camerlengo realizes that he has been discovered he sets himself on fire using holy oil from the Vatican and dies.
The Vatican announces to the media that the Camerlengo died due to internal wounds suffered during his landing and is canonized by the catholic church. The Conclave designate the surviving preferiti cardinal as the new Pope, and he gives Langdon the Galileo book that can help him finish his book in reward for saving his life before.
- Tom Hanks reprises his role as Professor Robert Langdon, a Harvard University professor of symbology.
- Ayelet Zurer plays Vittoria Vetra, a CERN scientist whose antimatter experiment has been stolen by the Illuminati to be used as a bomb.
- Ewan McGregor plays the Camerlengo Patrick McKenna (Carlo Ventresca in the book).
- Stellan Skarsgård plays Commander Richter, head of the Swiss Guard.
- Pierfrancesco Favino plays Inspector Ernesto Olivetti of the Gendarme Corps of Vatican City State.
- Nikolaj Lie Kaas plays Mr. Gray, an assassin who believes he is working for the Illuminati.
- Armin Mueller-Stahl plays Cardinal Strauss, elector of the conclave.
- Thure Lindhardt plays Lieutenant Chartrand, officer of the Swiss Guard.
Many characters have been changed for the film. These include:
- Vittoria Vetra's father, Leonardo Vetra, is replaced with a research partner named Silvano, who writes the journals and is murdered for his projects.
- The novel's Italian camerlengo, Carlo Ventresca, becomes the film's Northern Irish camerlengo, Patrick McKenna.
- The Swiss Guard commander, Commander Olivetti, becomes Commander Richter. Olivetti becomes the head of the Vatican police.
- The Hassassin, an Arabic man who hates the Catholic Church, is replaced with a Westerner named Mr. Gray.
- Captain Rocher does not appear in the film. Instead, his role near the book's climax is played by a bishop.
- Cardinal Mortati, who oversees the conclave, is renamed Cardinal Strauss.
- Gunther Glick and Chinita Macri do not appear in the film. Instead, numerous reporters from different countries perform reporting roles.
- Maximilian Köhler, director of CERN, does not appear in the film. Instead, his role in the beginning is played by a Vatican police representative, and his role near the end is played by Commander Richter.
Differences between book and film
- The funeral of the Pope is an original addition to the movie, as in the novel, the Pope is already buried and conclave is about to commence.
- In the novel, Langdon goes to Geneva after receiving a phone call from CERN. In the film, a representative of the Vatican Police visits him in Harvard, and tells him everything there.
- Vittoria is working on the antimatter at the beginning of the film with Silvano and a whole team, while in the novel, she returns to CERN after her father's murder. Also, only she and her father worked on the antimatter project.
- The Corps of Gendarmerie of Vatican City does not appear in the novel; the Swiss Guard handles all matters.
- The antimatter journals are sent over to the Vatican in the film, but Commander Richter confiscates them; this is how he learns of the truth.
- Vittoria is able to restore the antimatter canister's battery in the film by doing a few simple defusing processes to the canister as long as there is at least five minutes of time left. In the novel, she must fly the canister back to CERN to recharge it, so the canister is much more dangerous in the novel. However, in the movie she decides not to replace the battery since cold temperature in the Necropolis would be depleting the batteries faster; removing and replacing with new ones might not have been possible because the charge would not hold for sufficient time to replace the battery.
- In the novel, while confronting the assassin at the 3rd Altar of Science, Langdon survives by hiding under a stone tomb and is later found by police when his watch alarm goes off. In the film he hides down a grate in the church floor until the police and firefighters hear him banging.
- The Diagramma book written by Galileo Galilei during his house arrest is presented in the film as a simple booklet, while it is in folios in the novel. Hence, Vittoria permanently destroys it when she rips page five off in the film.
- Vittoria is not kidnapped during the Santa Maria della Vittoria incident; at that time, she is working on reading the antimatter journals.
- Langdon and Vittoria do not kill the assassin together. He is promised a reward for his actions but is blown up when a car bomb explodes in the car he was supposed to make his getaway in.
- The last brand in the novel is the Illuminati Diamond, which is a combination of "Earth", "Air", "Fire", and "Water" in the shape of an ambigram diamond. In the film, however, it is two keys crossed with each other, not even an ambigram. According to Langdon, its upside-down image is a reference to St. Peter's upside-down crucifixion.
- In the film, Langdon does not accompany the camerlengo in the helicopter, and thus does not injure himself in the Tiber River.
- In the film, Langdon and Vittoria discover the truth about the camerlengo in Commander Richter's office, as the Pope's seizures required security cameras to watch him. In the novel, Köhler gives him a small camera to watch the scene, while in the film, Richter gives him the key to the camera screen in his office.
- The truth about the Pope fathering the camerlengo is not mentioned in the film, although the camerlengo states the Pope adopted him. The only reason given for the camerlengo murdering him is that the Pope became too open to science, which the camerlengo was uncomfortable with, as he believed religion alone should dictate morality, let alone the Creation.
- The camerlengo immolates himself in the novel on the roof of St. Peter's, and dies calmly. In the film, he immolates himself using one of the ninety-nine oil lamps' oil as fuel, and the sanctuary lamp's flame as fire, and dies screaming.
- Cardinal Baggia, the fourth preferiti, survives his drowning in the film, as the assassin left Piazza Navona right away. Langdon manages to use the bubble-producing hose to save him, and manages to get bystanders to help rescue him. Baggia then informs Langdon that the four preferiti were imprisoned in Castel Sant'Angelo. As anticipated, he becomes Pope in the end, and Cardinal Strauss succeeds McKenna as the camerlengo instead. In the novel, Baggia also dies, and Mortati, despite his status as elector, becomes pontiff.
- The package loaned indefinitely to Langdon is not the last brand, but the Diagramma book written by Galileo Galilei.
- The love thread between Langdon and Vittoria in the book is not depicted in the film
- While the book Angels and Demons is a prequel to the Da Vinci Code, the movie is a sequel.
- The assassin gets inputs through internet in the film
- In the novel, Langdon goes alone to the archives the second time while looking for Bernini's statues. In the film, he is accompanied by a swiss guard
- The tourist guide in the Pantheon (novel) is replaced by a female guide
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