There is an urgent need to develop an efficient approach to limit biofilm formations. Recent progress in the biofilm treatment field showed remarkable utilization of emerging nanoformulations. In this study, we decorated clay halloysite nanotubules with tannic acid and Fe 3 O 4 Nanoparticles. The successful synthesis of the nanocomposite was validated by characterization techniques such as XRD, FE-SEM, HR-TEM, SEM-EDS, TEM-EDS, XPS, and VSM analysis. After successful characterizations of nanocomposite (HNTs-M-Tannic acid), antibiofilm effect analysis experiments such as Crystal Violet (CV) staining, biofilmforming marker gene expression analysis, morphology evaluation by SEM, and live and dead cell assay were performed. The Crystal Violet (CV) staining results concluded a significant 57.8% inhibition of the E. coli biofilm formation. Comprehensive gene expression analysis of f imA and csgA for E. coli biofilm formation confirmed the capacity of HNTs-M-Tannic acid to impede bacterial cell adhesion, thereby inhibiting biofilm formation. The SEM images and live and dead assay results confirmed and marked the potential of the developed HNTs-M-Tannic acid as an antibiofilm agent against E. coli. In conclusion, this study produced a unique nanocomposite, HNTs-M-Tannic acid, which offers potential as an economically viable, magnetically separable, and biocompatible nanoformulations for antibiofilm studies.