Background: Limited data are available on humoral responses determined by automated neutralization tests following the administration of the three different types of COVID-19 vaccinations. Thus, we here evaluated anti-SARS-CoV-2 neutralizing antibody titers via two different neutralization assays in comparison to total spike antibody levels. Methods: Healthy participants (n= 150) were enrolled into three subgroups who were tested 41 (22–65) days after their second dose of mRNA (BNT162b2/mRNA-1273), adenoviral vector (ChAdOx1/Gam-COVID-Vac) and inactivated whole-virus (BBIBP-CorV) vaccines, with no history or serologic evidence of prior SARS-CoV-2 infection. Neutralizing antibody (N-Ab) titers were analyzed on a Snibe Maglumi®800 instrument and a Medcaptain Immu F6®Analyzer in parallel to anti-SARS-CoV-2 S total antibody (S-Ab) levels (Roche Elecsys®e602). Results: Subjects who were administered mRNA vaccines demonstrated significantly higher SARS-CoV-2 N-Ab and S-Ab levels compared to those who received adenoviral vector and inactivated whole-virus vaccinations (p< 0.0001). N-Ab titers determined by the two methods correlated with each other (r = 0.9608;p< 0.0001) and S-Ab levels (r = 0.9432 and r = 0.9324;p< 0.0001, respectively). Based on N-Ab values, a new optimal threshold of Roche S-Ab was calculated (166 BAU/mL) for discrimination of seropositivity showing an AUC value of 0.975 (p< 0.0001). Low post-vaccination N-Ab levels (median value of 0.25 μg/mL or 7.28 AU/mL) were measured in those participants (n= 8) who were infected by SARS-CoV-2 within 6 months after immunizations. Conclusion: Both SARS-CoV-2 N-Ab automated assays are effective to evaluate humoral responses after various COVID-19 vaccines