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Around the same time, if not slightly later, there are intimations that Papias knew the gospels, perhaps even all four. The so-called "longer ending of Mark" (Mark 16:9-20), which most text critics regard as a spurious addition tacked on around A.D. 125, seems to reflect bits of Matthew, Luke, and John—and of course Mark itself. At the very least, this demonstrates that the four gospels were in broad circulation. It may even be the case, although it is impossible to prove, that the four gospels by this time constituted a collection in its own right, a sub-canon within the slowly emerging New Testament canon.

Firmer evidence for the four-fold gospel's authoritative status comes from the apologist Justin Martyr around the year 150. Following the philosophical terminology of his day, Justin preferred to call the gospels "memoirs." Justin records that the church used these "memoirs" regularly in their weekly services. This would seem to indicate that the four gospels had achieved a de facto canonical status.

Around this time the famed heretic Marcion of Sinope, convinced that it was necessary to eradicate any Jewish elements from the Christian Scriptures, took it upon himself to pare down the received collection of Christian books. The only gospel that "made the cut" was Luke, but only in an edited-down version. It has been widely argued (notably by Adolf von Harnack) that the Christians first began to think about the concept of a biblical canon only in response to Marcion. But given the evidence of Justin, this seems very unlikely. We are better off maintaining that it was actually Marcion who was reacting to an established de facto canon.

A fragmentary list of New Testament books known to us as the Muratorian Canon can, despite some scholarly opinion, be reasonably dated to around 170. This provides an important piece of second-century evidence for the gospels' canonicity. Unfortunately, the beginning of the Muratorian list is broken off. The fragment begins, "The third book of the gospel, according to Luke." It later continues, "The fourth gospel is by John." Interestingly, Luke is presented as the third gospel and John, the fourth—exactly the canonical order as we have it today. It is hardly a stretch to suppose that the missing first and second gospels were Matthew and Mark, respectively. Matthew, being the favorite gospel of the early church, was almost always positioned first in similar such lists and whenever the four-fold gospel was brought together in one volume.

The first church father to mention all four gospels by name was Irenaeus some time around 170. He strenuously objected to the various heretical sects (including that of Marcion) who had latched on to only one of the four gospels in order to substantiate their teachings. In Against Heresies, Irenaeus wrote:

The Gospels could not possibly be either more or less in number than they are. Since there are four zones of the world in which we live, and four principal winds, while the Church is spread over all the earth, and the pillar and foundation of the Church is the gospel, and the Spirit of life, it fittingly has four pillars, everywhere breathing out incorruption and revivifying men.

While this may not seem the most convincing line of argument today, Irenaeus's statement needs to be understood against the backdrop of a larger argument, which presupposed a theological correlation between creation (made up of four zones) and the new creation, Jesus Christ (revealed by four gospels). While certain scholars have accused Irenaeus of originating this policy of "these four and no more" in order to squelch competing sects with their gospels, the evidence for a much earlier four-fold gospel canon is more compelling.

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