In The Silence of the Lambs, the character of Clarice Starling is one of several whose perspectives we are privileged to explore. Other sections delve into the minds of Special Agent Jack Crawford, Hannibal Lecter, and Buffalo Bill/Jame Gumb. However, it is clear that Starling is the primary protagonist. The narrative begins and concludes with her, chronicling her mission to capture Buffalo Bill, with assistance from Dr. Lecter's clues. Clarice is intelligent, driven, and skilled: she graduated with honors from the University of Virginia, holding a double major in criminology and psychology. Though she is the daughter of a small-town lawman who was murdered by criminals, as Lecter notes, she is also motivated by compassion for the less fortunate.

Jame Gumb, also known as "Buffalo Bill" in the media, is the serial killer pursued by Clarice and law enforcement. Similar to the antagonist in Red Dragon (1981), his murders are part of his quest for transformation. However, Gumb's transformation is more straightforward: he is killing and skinning women to create a "woman suit" for himself. As a drifter with a poor self-image, Gumb desires to be anyone but himself. He is another of Harris's antagonists shaped by societal and psychological factors, yet Harris avoids simplistic explanations: "At least two scholarly journals explained that this unhappy childhood was the reason he killed women in his basement for their skins. The words crazy and evil do not appear in either article."

Hannibal Lecter plays a significantly more prominent and active role in this novel compared to Red Dragon. His interactions with Starling form the heart of the story, and through their exchanges, he comes to respect and even develop a liking for her. This glimmer of humanity makes Lecter captivating, as unlike Gumb, no effort is made to justify his transformation into what he has become. He is depicted as purely evil, and his escape from captivity in a violent yet ingenious manner serves as a reminder of his intricate personality—a man of sophistication and elegance who has killed and consumed at least nine individuals.