IntroductionHepatitis B virus infection is still an important health problem worldwide. The risk of contracting HBV by health-care workers (HCWs) is higher than that of the general adult population. Seroprotection is defined as anti-HBs level 10mIU/ml or more following a successful three-dose vaccination course. ObjectiveTo find out the hepatitis B vaccination status and test immune response using anti-HBs titer among HCWs. MethodsWe carried out a retrospective observational study of 218 serum samples from vaccinated HCWs tested for anti-HBs ELISA from January 2016 to December 2017. We collected demographic, health, and HBV vaccination details from test requisition forms. ResultsWe found that 195 (89.44%) HCWs had received a complete course of HBV vaccination. 74.87% HCWs showed a protective level (10mIU/ml or more) of antibodies. We noted a statistically significant association between the protective antibody level and the complete course of vaccination (P<0.001). HCWs had protective levels of antibodies even after 25 years post-vaccination. We found a statistically significant decrease in mean antibody titer with duration (in years) of post-primary vaccination (P<0.001). We noted higher seroprotection in females, age group of 21–30 years, and weight range of 51–70kg. ConclusionHepatitis B vaccination coverage is still inadequate even among HCWs. Even with recommended three dose full vaccination, some HCWs have not achieved protective anti-HBs titer, so we have to make testing of anti-HBs compulsory irrespective of the vaccination status. We noted a decrease in mean antibody titer with duration (in years) of post-primary vaccination.