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Susan Cooper (Melissa McCarthy) is a 40-year-old, single, desk-bound CIA employee who remotely assists her partner, field agent Bradley Fine (Jude Law), on a mission. Fine accidentally kills arms dealer Tihomir Boyanov before extracting the location of a suitcase nuke from him. Susan uncovers evidence that Rayna (Rose Byrne), Boyanov's daughter, has contacted Sergio De Luca (Bobby Cannavale), a suspected broker with ties to various terrorist groups, so Fine infiltrates her home. However, Rayna shoots Fine dead, while Susan watches helplessly online, then reveals that she knows the identities of the agency's top agents, including Rick Ford (Jason Statham) and Karen Walker (Morena Baccarin). Susan, who is almost certainly unknown to Rayna, volunteers to track him. (She was a top trainee agent, albeit over ten years ago.) When her boss, Elaine Crocker (Allison Janney), reluctantly agrees, the ultra macho Ford quits in disgust.
With her best friend Nancy (Miranda Hart) providing intelligence, Susan goes to Paris undercover. That night, Ford shows up and tells Susan she will fail because of her inexperience. The next morning, Susan discovers that De Luca's office has burned down. She finds a photo of a man standing next to the fire. Ford appears, argues with Susan again and leaves. Susan sees the man in the photo follow him and switch his backpack with another one containing a bomb while Ford is distracted. Susan warns Ford in time and pursues the man into an abandoned building. During the ensuing fight, he falls to his death. When she checks the man's video camera, Susan learns that De Luca is going to Rome.
In Rome, Susan meets her contact Aldo (Peter Serafinowicz). She follows Sergio into a casino, where she saves Rayna's life. Rayna brings Susan into her inner circle and takes Susan on her private plane to Budapest. In mid-flight, the steward kills Rayna's bodyguard and pilots, but Susan subdues him. Rayna believes Susan to be a CIA agent, but Susan convinces her that she was hired by Boyanov to protect his daughter.
In Budapest, Susan meets Nancy, who was sent by Crocker. After being shot at, Susan pursues and catches up with the would-be assassin: Karen, who sold Rayna the names of the other agents. She tries to shoot Susan, but an unknown sniper kills her. Susan, Nancy and Aldo accompany Rayna to a party to meet Rayna's contact. That turns out to be Lia (Nargis Fakhri), the woman who distracted Ford in Paris. Nancy creates a diversion (by pretending to be a crazed fan of guest performer 50 Cent) so that Susan can try to apprehend Lia unnoticed. Because of Ford's inopportune intervention, however, Lia runs off. Susan chases after her. After a brutal fight, Lia is about to eliminate Susan, but is herself killed by Fine, who earlier faked his death and is now Rayna's lover and associate.
Rayna imprisons Susan and Aldo in a bunker. Later, Fine reveals to Susan that he is trying to gain Rayna's trust to locate the nuke, and he was the one who killed Karen. Susan and Aldo escape. Susan convinces Rayna that, even though she works for the CIA, she will do anything to protect Fine, admitting that she loves him.
At De Luca's mansion, Fine, Rayna and Sergio wait for Solsa Dudaev (Richard Brake), head of an al-Qaeda-funded terrorist group. Dudaev gives De Luca a suitcase full of diamonds, and Rayna produces the device. De Luca has Dudaev and his men killed, then reveals his plan to sell the device to another buyer (though they also intend to bomb New York City), before pointing his gun at Rayna. Ford distracts him, allowing Susan to disarm him. Sergio escapes to his helicopter with the device and the diamonds, but Susan grabs onto the landing gear. In the ensuing struggle, the diamonds and the device fall into the lake. Nancy follows in another helicopter and shoots Sergio before he can kill Susan. Rayna is arrested and the nuke retrieved. Ford, to Susan's surprise, compliments her on her job. Fine invites Susan to dinner, but she instead opts for a night out with Nancy.
The next morning, Susan wakes up in bed next to Ford and screams. It is implied that they slept together, and Ford says that she loved it.