A novel and ideal immunosensor platform with customized tailored chemistry is suggested by combining the characteristics of nanofibers (NFs) and dendrimers. Polystyrene (PS) NFs were doped with polyamidoamine (PAMAM) dendritic polymer and utilized for covalent immobilization of FITC-labeled anti-human TNF-alpha antibodies by activation of the amine groups of dendrimer on the surface of NFs using a glutaraldehyde (GA) coupling agent. SEM, ATR-FTIR, contact angle, EDX, XPS, TGA, and fluorescence microscopy were performed. The findings demonstrated that (1) PS-PAMAM NFs were produced with significantly smaller diameters (295 +/- 94 nm), more uniform morphology, and more hydrophilic (44 +/- 5.5 degrees) with dendrimer amine groups on the surface that can provide a better path for antibodies to be immobilized compared to pure PS NFs, (2) the PAMAM was distributed evenly throughout the NFs, (3) GA was uniformly applied to the surface of NFs via covalent bonding, (4) the PAMAM played a critical role in substantially increasing antibody immobilization at the surface of each NFs without causing agglomeration. According to the findings, the P5-PAMAM electrospun mat is suitable for immobilizing biomolecules and may be utilized as a support to boost immunosensor sensitivity.