Peptide-based nanomaterials can be easily functionalized due to their functional groups, as well as being biocompatible, stable under physiological conditions, and nontoxic. Here, diphenylalanineamide-based nanomaterials (FFANMs) were synthesized, decorated with Ca ions to set the surface charge, and characterized for possible use in gene delivery and drug release studies. FFANMs were characterized by SEM, TEM, dynamic light scattering (DLS), and LC-MS/MS. Corona formation and biocompatible studies were also carried out. Some of the data obtained are as follows: FFANMs have a diameter of approximately 87.93 nm. While the zeta potentials of FFANMs and Ca@FFANMs were -20.1 mV and +9.3 mV, respectively, after corona formation with HSA and IgG proteins, they were shifted to -7.6 mV and -3.7 mV, respectively. For gene delivery studies, zeta potentials of Ca@FFANMs and DNA interactions were also studied and found to shift to -9.7 mV. Cytotoxicity and biocompatibility studies of NMs were also studied on HeLa and HT29 cell lines, and decreases of about 5% and 10% in viability at the end of 24 h and 72 h incubation times were found. We think that the results obtained from this study will assist the groups working in the relevant field.