This study was intended to investigate the crosslinking characteristics of a new crosslinking agent-oxidized sodium alginate (ADA), which might provide an ideal biological crosslinking reagent for the construction of soft tissue bioprostheses. Glutaraldehyde and genipin, which have been typically used in developing bioprostheses, were used as controls. The porcine aortas were treated by these three crosslinking agents for 15 min to 72 h and the fixation index was determined. Subsequently, the mechanical property and cytocompatibility of fixed tissues were also tested. The results indicated that fixed tissues by ADA were comparable as glutaraldehyde and superior to genipin controls in fixative efficiency. It was also found that tissues fixed by ADA were comparable as genipin and superior to glutaraldehyde controls in cytocompatibility and were similar to natural tissues in mechanical property. The results of in vitro study demonstrated that ADA could be a promising crosslinking reagent for biological tissue fixation.