The effective functionalization and self-assembly of MXene are of crucial importance for a broad range of nanomaterial applications. In this work, we investigated the aggregates of sulfanilic acid-modified MXene (abbreviated as MXene-SO3H) with three model dyes at the air–water interface and demonstrated the morphological and aggregation changes of composite films, using Langmuir-Blodgett (LB) technology, as well as excellent uniformity and reproducibility by using surface-enhanced Raman scattering (SERS) spectra. This research has found that cationic dye molecules were adsorbed onto negatively charged MXene-SO3H particles mainly through electrostatic interaction and the particles induced dyes to form highly ordered nanostructures including H- and/or J-aggregates corresponding to monomers in bulk solution. The surface pressure-area isotherms from different dye sub phases confirmed that the stable composite films have been successfully formed. And the spectral results reveal that different dyes have different types of aggregations. In addition, the SERS spectra indicated that the optimal layers of MXene-SO3H/methylene blue (MB) films was 50 layers using rhodamine 6G (R6G) as probe molecule. And the formed 50 layers of MXene-SO3H/MB films (MXene-SO3H/MB-50) as SERS substrate were proved to possess excellent uniformity and repeatability.