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The development and spread of bacterial resistance to antimicrobial drugs necessitates the need to search for novel and effective antimicrobial agents. In the last few decades, innovative nanomaterials are attracting increasing attention and, among them, dendrimers have shown wide application in the various fields. In the current study, the two generations of an anionic linear- spherical nanodendrimer G1 and G2 were synthetized and compound G2 of nanodendrimer conjugated with erythromycin. The structures of the nanodendrimers were characterized by FTIR spectroscopy, zetasizer, and scanning electron microscopy (SEM). The antibacterial activity of the erythromycin-conjugated nanodendrimer and erythromycin alone were evaluated by the microdilution method against Staphylococcus aureus, S. epidermidis, S. saprophyticus, and Pseudomonas aeruginosa. The size of first and second generation of nanodendrimer, and the erythromycin-conjugated nanodendrimer was 75, 95, and 65.6 nm, respectively. The drug loading percentage of the nanodendrimer conjugates was obtained to be in 35.2%. In our study, the erythromycin-conjugated nanodendrimer showed significantly more bacteriostatic and bactericidal activities against all four studied bacteria than erythromycin alone. Our study's results highlight that the erythromycin-conjugated nanodendrimer is a highly effective agent against Gram positive and negative bacteria. The antibacterial properties of erythromycin combined with the targeting potential of the nanodendrimer can lead to sustained intracellular delivery of therapeutic agent.