We recently developed a novel biological glue from tartaric acid derivative (TAD) with two active ester groups and human serum albumin (HSA), named TAD-A. In this study, in vivo experiments were performed to investigate clinical applicability of TAD-A. TAD was prepared by reacting carboxyl groups of tartaric acid with N-hydroxysuccinimide in the presence of carbodiimide. Bonding strength was evaluated by using mouse skin closed with TAD-A of different TAD concentrations from 0.1 to 0.5 mmol in 0.8 mg of 44 w/w % HSA solution. Commercially available glues such as fibrin and aldehyde-based glue were used for comparison. We found that TAD-A's bonding strength increased significantly with TAD-A concentration. The bonding strength of 0.5 mmol of TAD-A in 0.8 mg of 44 w/w % HSA solution was significantly higher than that of fibrin or aldehyde-based glue (p < 0.01), and that of 0.3 mmol of TAD-A was significantly higher than of fibrin glue (p < 0.05). To determine toxicity, we implanted disks made from TAD-A of different TAD concentrations from 0.1 to 0.5 mmol in 0.8 mg of 44 w/w % HSA solution subcutaneously in mice. The inflammatory reaction in surrounding tissue increased with increasing TAD concentration, and then the disks were absorbed. In conclusion, TAD-A has sufficient bonding strength and comparatively low toxicity in clinical use of 0.3 mmol or less of TAD and 0.8 mL of 44 w/w % HSA solution.