Canine parvovirus-2 (CPV-2) infection causes serious multisystemic disease in dogs and many animal species worldwide. Previously, a monoclonal antibody (MAb) of CPV-2, 10H4, showed high neutralizing activity and therapeutic effect against CPV-2 in dogs. However, the application of mouse MAb is limited in other animals due to immune rejection. Here, the variable regions of the heavy and light chains of 10H4 were cloned and ligated with constant canine antibody regions to produce a canine-derived chimeric MAb 11D9, in a CHO-S cell expression system. The cell supernatant of the CHO cell line 11D9 exhibited a HI titer of 1:2560 against all the variants of CPV-2 (new CPV-2a, new CPV-2b, and CPV-2c), and had the same average neutralization titer as the new CPV-2a (1:11,046.5) and new CPV-2b (1:11,046.5) variants, which was slightly higher than that of CPV-2c variants (1:10,615.7). In animal experiment, the treatment of chimeric MAb 11D9 had a high therapeutic effect in beagles infected with the new CPV-2a. Overall, the canine-derived chimeric MAb 11D9 produced by CHO-S cells showed a high HI and neutralization titer against CPV-2 and the therapeutic effects against the new CPV-2a in beagles, providing potential for the prevention or treatment of CPV-2 infections in dogs.