Two-dimensional covalent organic frameworks (COFs) were used to create the first asymmetric, thin-film composite (TFC) nanofiltration (NF) membrane with a COF active layer. NF membrane active layers of polyimine COF were synthesized via the interfacial polymerization (IP) of terephthalaldehyde and tris(4-aminophenyl)benzene monomers on top of a poly(ether sulfone) (PES) ultrafiltration membrane support. Rutherford backscattering spectrometry and Fourier transform infrared spectroscopy analyses confirmed the presence of an imine-linked film with a thickness of ∼10 nm that was formed reproducibly. The rejection efficiencies of the COF NF membrane for a model organic compound, Rhodamine-WT, and a background electrolyte, NaCl, were higher than those of the PES support without the COF film. This enhanced solute rejection is the first successful demonstration of a TFC membrane with a thin COF active layer. However, this work also demonstrates the need for COF NF membranes with smaller active layer pores and alternative support materials. The former should result in greater solute rejection, and the latter is key because the PES used for support in the COF membranes is incompatible with the organic solvents used for the COF IP process.