The novel thin film composite (TFC) forward osmosis (FO) membrane with electrospinning nanofibers as support layer can alleviate internal concentration polarization (ICP). While the macropores of the nanofiber support layer cause defects in the polyamide (PA) layer. Therefore, hydrophobic polyvinylidene fluoride (PVDF) fine nanofibers were used as an interlayer to modulate the process of interfacial polymerization (IP) in this study. The results showed that the introduction of the interlayer improved the hydrophobicity of the support layer for achieving uniform, thin and defect-free selective polyamide (PA) layer. The water flux of TFC-PVDF was 58.26 LMH in the FO mode of 2 M NaCl, which was two times higher than that of the unmodified FO membrane. Lower reverse salt flux (4.91 gMH) and structural parameter (179.43 μm) alleviated the ICP. In addition, TFC-PVDF membrane showed good anti-fouling performance for SA (flux recovery ratio of 93.97%) due to high hydrophilicity, low zeta potential and low roughness. This study provides an easy and promising method to prepare defect-free PA selective layer on the macropores nanofiber support layer. The novel FO membrane shows high desalination performance and anti-fouling properties.