To improve the adsorption efficiency of pollutants by biochar, preparing graphene-like biochar (GBC) or nitrogen-doped biochar are two commonly used methods. However, the difference in the nitrogen doping (N-doping) effects upon the adsorption of pollutants by pristine biochar (PBC) and GBC, as well as the underlying mechanisms, are still unclear. Take the tetracycline (TC) as an example, the present study analyzed the characteristics of the adsorption of TCs on biochars (PBC, GBC, N-PBC, N-GBC), and significant differences in the effects of N-doping on the adsorption of TCs by PBC and GBC were consistently observed at different solution properties. Specifically, N-doping had varied effects on the adsorption performance of PBC, whereas it uniformly improved the adsorption performance of GBC. To interpret the phenomenon, the N-doping upon the adsorption was revealed by the QSAR model, which indicated that the pore filling (V_M) and the interactions between TCs with biochars (E_ad-v) were found to be the most important two factors. Furthermore, the density functional theory (DFT) results demonstrated that N-doping slightly affects biochar’s chemical reactivity. The van der Waals (vdWs) and electrostatic interactions are the main forces for TCs-biochars interactions. Moreover, N-doping mostly strengthened the electrostatic interactions of TCs-biochars, but the vdWs interactions of most samples remained largely unaffected. Overall, the revealed mechanism of N-doping on TCs adsorption by biochars will enhance our knowledge of antibiotic pollution remediation.