A mysterious aircraft steals a highly potent mutagen known as PX-41 from a laboratory in the Arctic Circle. Silas Ramsbottom, director of the Anti-Villain League (AVL), sends agent Lucy Wilde to recruit former supervillain Gru, who is forcibly brought to their headquarters and asked to help them track down the culprit and recover the mutagen. However, Gru refuses and informs Silas and Lucy that he has enough responsibilities as a father and as the owner of a jelly-producing business. Later, Dr. Nefario, Gru's assistant, quits to accept a job offer to assist another villain; this causes a reluctant Gru to decide to work with the AVL after all. He and Lucy are partnered and stationed at a shopping mall, with a cupcake store as their front.