In recent years dendrimers have fascinated the investigators towards targeted drug delivery because of their versatile framework and exhibit immense potentiality in entrapping drug moieties through host-guest interactions and serve as a promising vector in biological applications. The current investigation is focused on developing pegylated citric acid cefotaxime dendrimers through the divergent method and its characterization through spectroscopic, microscopic, thermal and microscopic techniques. Among the spectroscopic techniques, 1H NMR and 13C NMR elucidated the key functional groups at various chemical shifts while ESI-MS pointed out the molecular weight of cefotaxime sodium in various generations. Similarly, FTIR, DSC, and AFM investigations detailed that the generations are devoid of incompatibilities, structural deformities and can be opted for targeted drug delivery. The drug entrapment studies and in-vitro drug release studies highlight CFTX G5 containing 92.4% entrapment efficacy and 83.8% drug release in 48 h and specifies a sustain release characteristics. In connection to the above, the in-vivo studies reveal a potent antibacterial activity against various gram-positive and gram-negative microorganisms with a decreased hemolysis and cytotoxicity effects and reflect a high margin of safety regarding pegylated CFTX dendrimers. Further, the antibacterial activities are supported through confocal microscopy that clarified the cellular uptake of dendritic molecules and their internalization. © 2019 Elsevier B.V.