- Agnes is Don Lorenzo's younger sister and Don Raymond's lover. Her mother fell ill while pregnant with Agnes and vowed to send Agnes to the convent if she delivered her safely. She is a virtuous young lady who intends to marry Don Raymond but her parents want her to become a nun, so she decides to run away with him. Their plans are foiled and, thinking Don Raymond has abandoned her forever, she enters the convent.
- Ambrosio is an extremely devout monk about 30 years old. He was found left at the Abbey doorstep when he was too young to tell his tale. The monks consider him a present from the Virgin Mary and they educate him at the monastery.[2]
- Antonia is a timid and innocent girl of 15. She was brought up in an old castle in Murcia with only her mother Elvira and is therefore very sheltered. She is the object of Don Lorenzo's attentions. The novel's evil characters are considered to be better written than the virtuous ones, and Antonia's character is so virtuous that some have found her "deadly dull".[3]
- Elvira is the mother of Antonia and Ambrosio. She married a young nobleman in secret. His family does not approve of her and because of this she and her husband escape to the Indies, leaving her 2-year-old son behind. After 13 years, when Antonia is very young, her husband dies and she returns to Murcia where she lives on an allowance given to her by her father-in-law.
- Leonella is Elvira's sister and Antonia's spinster aunt. She takes an immediate dislike to Ambrosio after hearing his sermon.[4] She believes Don Christoval's polite attentions are more significant than they actually are and is hurt when he fails to call at her house. She eventually marries a younger man and lives in Cordova.
- Don Lorenzo de Medina is Agnes's older brother and friend of Don Raymond and Don Christoval. Immediately intrigued by Antonia after meeting her at Ambrosio's sermon, Don Lorenzo resolves to marry her.
- Matilda is first known as Rosario, the young boy who looks up to Ambrosio "with a respect approaching idolatry".[5] Rosario is brought to the Monastery by a well dressed rich stranger but not much more is known of his past. He always hides under his cowl and later reveals that he is actually Matilda, a beautiful young lady who loves Ambrosio. Matilda 'loved' Ambrosio even before she joined the monastery (as a boy), and therefore requested a painting of herself as Madonna to be given to Ambrosio, which hangs in his room. She seduces Ambrosio and aids in his destruction of Antonia with magic. The character of Matilda was highly praised by Coleridge as Lewis's masterpiece, and is said to be "exquisitely imagined"[6] and "superior in wickedness to the most wicked of men."[7] Though she is considered by some critics to be the most intelligent, articulate, and interesting,[8] she is difficult to characterise. The plot of the novel relies on her being a supernatural force with magical powers,[9] but she begins as a human. She tells Ambrosio she loves him when she thinks he is asleep and cries "involuntary" tears when she realises he no longer cares for her. These passages, together with the haste in which the novel was written, seem to indicate "that Lewis changed his mind in the course of the narrative".[10]
- The Prioress, also known as Mother St. Agatha, punishes Agnes severely to uphold the honour of the convent of St. Clare. "Viciously cruel in the name of virtue",[3] she keeps Agnes prisoner in the dungeons beneath the convent with only enough bread and water to sustain her but not nourish her. The prioress circulates the story of Agnes's death to everyone, including Agnes's own relations. She is beaten to a bloody pulp by the crowd that gathers to honour St. Clare when they realise she is responsible for Agnes's supposed death. She is also the inspiration for the Abbess of San Stephano in Radcliffe's The Italian.[11]
- Don Raymond is the son of a Marquis and is also known as Alphonso d'Alvarada. He takes the name Alphonso when his father, by the urging of his friend the Duke of Villa Hermosa, advises him that taking a new name will allow him to be known for his merits rather than his rank. He travels to Paris, but finds the Parisians "frivolous, unfeeling and insincere"[12] and sets out for Germany. Near Strasbourg he is forced to seek accommodations in a cottage after his chaise supposedly breaks down. He is the target of the robber Baptiste but with help from Baptiste's wife Marguerite, Raymond is able to save himself and the Baroness Lindenberg. Grateful, the Baroness invites Don Raymond to stay with her and her husband at their castle in Bavaria.
- Donna Rodolpha, Baroness of Lindenburg meets Don Raymond while travelling to Strasbourg. She is in love with Don Raymond and becomes jealous when she finds out Don Raymond is in love with her niece, Agnes. She asks him to leave the Castle of Lindenberg and later speaks poorly of his character.
- Mother St. Ursula assists in Agnes's rescue. She is a witness to the Prioress's crimes and without her, Don Lorenzo would not be able to accuse the Prioress.
- Theodore is Don Raymond's page. He enjoys writing poetry and authors the poems "Love and Age" and "The Water King". After reading "Love and Age", Don Raymond points out the flaws in the piece, which may be flaws Lewis noticed in his own work.[10] Far from being the unwaiveringly faithful servant stock character,[13] Theodore plays a key role in moving the plot forward by helping with Don Raymond's plans to escape with Agnes. Theodore's character also provides foreshadowing through his poems. His poems parallel the action of the story. For instance, in his poem "The Water King", the lovely maid's fate foreshadows Antonia's.[14] In addition, Theodore also bears a striking resemblance to other characters in other of Lewis's works, including Leolyn in One O'clock (1811) and Eugene in "Mistrust" from Romantic Tales (1808).[15]
- Virginia de Villa Franca, introduced late in the story, is a beautiful, virtuous young relation of the Prioress who represents St. Clare in the Procession. Virginia nurses the ill Agnes back to health and thus wins Lorenzo's affections. Like Isabella in The Castle of Otranto, she is introduced as an acceptable marriage partner for Lorenzo, but plays an unessential part in the plot.[3]
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