Prosthetic vascular graft infection is one of the most severe vascular surgery complications. Fibrin gel (FG) has many useful characteristics as biocompatibility, biodegradation, adhesion, and haemostasis to develop the local antibiotic delivery system. In this study, human plasma was collected from peripheral blood that was used to create fibrin gel by supplement ion Ca2+. Antibiotic-containing fibrin gel was then evaluated in some characteristics such as surface structure, biodegradation, antibiotic delivery, cytotoxicity, and bacterial biofilm prevention in vitro and in vivo. The results showed that fibrin gel was excellent material for the extended delivery of antibiotics. Most importantly, antibiotic-containing fibrin gel was not toxic for human fibroblast cells in vitro and inhibited bacterial biofilm growth in vitro and in vivo. This research is the first step in developing an antibiotic delivery system for effective graft infection treatments.