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