question: (1) Background: Healthcare workers (HCWs) are prone to intensified exposure to severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection in the ongoing pandemic We prospectively analyzed the prevalence of antibodies against SARS-CoV-2 in HCWs at baseline and follow up with regard to clinical signs and symptoms in two university hospitals in Brandenburg, Germany (2) Methods: Screening for anti-SARS-CoV-2 IgA and IgG antibodies was offered to HCWs at baseline and follow up two months thereafter in two hospitals of Brandenburg Medical School during the first wave of the COVID-19 pandemic in Germany in an ongoing observational cohort study Medical history and signs and symptoms were recorded by questionnaires and analyzed (3) Results: Baseline seroprevalence of anti-SARS-CoV-2 IgA was 11 7% and increased to 15% at follow up, whereas IgG seropositivity was 2 1% at baseline and 2 2% at follow up The rate of asymptomatic seropositive cases was 39 5% Symptoms were not associated with general seropositivity for anti-SARS-CoV-2;however, class switch from IgA to IgG was associated with increased symptom burden (4) Conclusions: The seroprevalence of antibodies against SARS-CoV-2 was low in HCWs but higher compared to population data and increased over time Screening for antibodies detected a significant proportion of seropositive participants cases without symptoms