Background Estimates from initial SARS-CoV-2 serological surveys were likely to be biased due to convenience sampling whereas large-scale population-based serosurveys could be biased due to non-response. This study aims to estimate the seroprevalence of SARS-CoV-2 infection in Saint Petersburg, Russia accounting for non-response bias. Methods We recruited a random sample of adults residing in St. Petersburg with random digit dialling. Computer-assisted telephone interview was followed by an invitation for an antibody test with randomized rewards for participation. Blood samples collected between May 27, 2020 and June 26, 2020 were assessed for anti-SARS-CoV-2 antibodies using two tests — CMIA and ELISA. The seroprevalence estimates were corrected for non-response bias, test sensitivity, and specificity. Individual characteristics associated with seropositivity were assessed. Findings 66,250 individuals were contacted, 6,440 adults agreed to be interviewed and were invited to participate in the serosurvey. Blood samples were obtained from 1038 participants. Naive seroprevalence corrected for test characteristics was 9.0% [95% CI 7.2–10.8] by CMIA and 10.8% [8.8–12.7] by ELISA. Correction for non-response bias decreased seroprevalence estimates to 7.4% [5.7–9.2] for CMIA and to 9.3% [7.4–11.2] for ELISA. The most pronounced decrease in non-response bias-corrected seroprevalence was attributed to the history of any illnesses in the past 3 months and COVID-19 testing. Besides that seroconversion was negatively associated with smoking status, self-reported history of allergies and changes in hand-washing habits. Interpretation These results suggest that even low estimates of seroprevalence in Europe’s fourth-largest city can be an overestimation in the presence of non-response bias. Serosurvey design should attempt to identify characteristics that are associated both with participation and seropositivity. Further population-based studies are required to explain the lower seroprevalence in smokers and participant reporting allergies. Funding Polymetal International plc