Nearly 80% of human chronic infections are caused due to bacterial biofilm formation. The increased resistance against the conventional antimicrobial agents makes it difficult to treat the biofilm-related infections. The antibiotics resistance developed by planktonic cells has also become a major threat for human. Therefore, we have attempted here to develop an effective alternative strategy to overcome the issues of antibiotics resistance of bacteria. Upon synthesis, biogenic C-dots were combined with lysozymes which were further encapsulated into chitosan nanocarrier to form C-dots carrier (CDC). The as-synthesized C-dots were found irregular shaped and the average size of C-dots and CDC were 8 ± 2 nm and 450 ± 50 nm, respectively. To ensure secure and targeted delivery of C-dots and lysozyme we have employed chitosan, a biodegradable and natural biopolymer, as a delivery system. The study of time-dependent bacterial growth and flow cytometry analysis demonstrated that CDC can exhibit a synergistic bactericidal activity against the antibiotics resistant recombinant E. coli cells. Further, we have shown that the CDC could be a potent agent for both prevention of biofilm formation and eradication of preformed biofilm. In addition, we have observed that our drug delivery system is hemocompatible in nature making it suitable for in vivo applications. Therefore, we believe that the combination therapy of C-dots and lysozyme may be used as an excellent antibacterial and antibiofilm strategy.