An enzyme-linked immunosorbent assay for anthrax antibody in white-tailed deer (Odocoileus virginianus) was developed and used to evaluate a vaccination study and compare sera from hunter-killed deer in anthrax endemic and non-endemic areas. Deer subcutaneously vaccinated with anthrax avirulent spore vaccine developed specific antibody to protective antigen (PA) which was significantly higher than the non-vaccinated controls at 30, 60, 90, and 240 days post-vaccination. There was no difference between the levels of antibody to PA between deer in anthrax endemic and non-endemic areas.