AbstractAutoantibodies against angiotensin‐converting enzyme 2 (ACE2) are frequently reported in patients during coronavirus disease 2019 (COVID‐19) with evidence for a pathogenic role in severe infection. However, little is known of the prevalence or clinical significance of ACE2 autoantibodies in late convalescence or following COVID‐19 vaccination. In this study, we measured ACE2 autoantibodies in a cohort of 182 COVID‐19 convalescent patients, 186 COVID‐19 vaccine recipients, and 43 adolescents with post‐mRNA vaccine myopericarditis using two ACE2 enzymatic immunoassays (EIAs). ACE2 IgM autoantibody EIA median optical densities (ODs) were lower in convalescent patients than pre‐COVID‐19 control samples with only 2/182 (1.1%) convalescents testing positive. Similarly, only 3/182 (1.6%) convalescent patients tested positive for ACE2 IgG, but patients with history of moderate‐severe COVID‐19 tended to have significantly higher median ODs than controls and mild COVID‐19 patients. In contrast, ACE2 IgG antibodies were detected in 10/186 (5.4%) COVID‐19 vaccine recipients after two doses of vaccination. Median ACE2 IgG EIA ODs of vaccine recipients were higher than controls irrespective of the vaccine platform used (inactivated or mRNA). ACE2 IgG ODs were not correlated with surrogate neutralizing antibody levels in vaccine recipients. ACE2 IgG levels peaked at day 56 post‐first dose and declined within 12 months to baseline levels in vaccine recipients. Presence of ACE2 antibodies was not associated with adverse events following immunization including myopericarditis. One convalescent patient with ACE2 IgG developed Guillain−Barre syndrome, but causality was not established. ACE2 autoantibodies are observed in COVID‐19 vaccine recipients and convalescent patients, but are likely innocuous.