Lotto kept a detailed account book, which on 8 December 1540 mentions the arrival in his studio of a canvas already nailed to a frame.

From this fact, art historians have hypothesized that the commission for the Antoninus was originally signed around 1525 and was the reason for Lotto's move from Bergamo to Venice, where he stayed in the Dominican monastery.

His work on the painting may have been interrupted in July 1526, when the artist left the monastery in disgust at certain slanders circulating about him.

The work went wholly unmentioned in Venice at the time of its production due to the intelligentsia's hostility to Lotto and preference for Titian and his followers.

In his composition, Lotto creates a model for bureaucratic charity—with the saint scrupulously reading requests, receiving guidance from the angels at his ears, and then giving instructions to those below.