Colantonio flourished in Naples between 1440 and 1470. He was a student of René d'Anjou. His paintings show the mingling of several cultures, as Alfonso V of Aragon had brought to Naples artists from Iberia, Burgundy, Provence, and Flanders. He synthesised his own style from all these sources and he was one of the first artists in Southern Italy to learn Flemish oil painting techniques. He may have learnt these techniques from the Flemish artist Bartélemy d'Eyck—a putative relative of Jan van Eyck—who seems to have been in Naples around 1440.
His most notable work is St. Francis consigns the Rule to his followers (dated to 1445). This painting—for the altarpiece of a transept chapel—was inspired by a late thirteenth century fresco above the door of the Chapter House of San Lorenzo Maggiore in Naples, showing the saint giving his Rule to the Minor Friars and the Poor Clares. The fresco can be seen in the monastery Museum. The painting hangs in the Museo di Capodimonte in Naples with other examples of his work.