2-Dodecyldiethylenetriaminepentaacetic acid (C12-DTPA) is a chelating, amphoteric surfactant with a bulky headgroup containing eight pH-responsive groups. The hypothesis was that the amphoteric nature of the chelating surfactant would affect the interaction with another surfactant and, consequently, also the composition of mixed surface layers. Binary mixed monolayers of C12-DTPA and the anionic surfactant sodium dodecyl sulfate (SDS) were examined using neutron reflection and surface tension measurements. The experiments were conducted at pH 5, where the C12-DTPA monomers carried a net negative charge. Surface excess calculations at low total surfactant concentration revealed that the chelating surfactant dominated the surface composition. However, as the concentration was raised, the surface composition shifted toward an SDS-dominant state. This phenomenon was attributed to the increased ionic strength at increased concentrations, which altered the balance between competing entropic forces in the system. Interaction parameters for mixed monolayer formation were calculated, following a framework based on regular solution theory. In accordance with the hypothesis, the chelating surfactant’s ability to modulate its charge and mitigate repulsive interactions in the surface layer resulted in favorable interactions between the anionic SDS and negatively charged C12-DTPA monomers. These interactions were found to be concentration-dependent, which was consistent with the observed shift in the surface layer composition.