We describe validation of a COVID-19 antibody test for detection of anti-SARS-CoV-2 receptor-binding domain (RBD) IgG antibodies in blood plasma utilizing ethically sourced reagents not derived from aborted fetal cell lines. The test demonstrated specificity of 100% (95% confidence intervals 77.2-100%) and sensitivity of 100% (95% confidence intervals 79.6-100%) when evaluating blood specimens previously determined to be negative (n = 13) or positive for anti-SARS-CoV-2 RBD IgG antibodies due to natural SARS-CoV-2 exposure (n = 13) or COVID-19 vaccination (n = 15). The test was used to screen 230 blood specimens from individuals with unknown SARS-CoV-2 exposure (n = 103) or that were naturally exposed to SARS-CoV-2 (n = 44), received a COVID-19 vaccine (n = 66), or received a COVID-19 vaccine before or after SARS-CoV-2 exposure (n = 17). Ninety-nine percent (95% confidence intervals 95.7-100%) of the 127 blood specimens from individuals that were naturally exposed, vaccinated, or both vaccinated and naturally exposed were positive for anti-SARS-CoV-2 RBD IgG which was consistent with the high sensitivity of our test. This COVID-19 antibody test, now named the PL COVID-19 RBD IgG antibody test, represents an effective and ethical alternative to commercially available COVID-19 antibody tests that utilize reagents derived from aborted fetal lines.