A new paper-based lateral flow immunoassay configuration was engineered and investigated. The assay is intended for the detection of a model protein in human serum, that is, human immunoglobulin G, with the aim to demonstrate a virtually universal protein detection platform. Once the sample is added in the strip, the analyte is selectively captured by antibody-decorated silica beads (Ab-SiO2) onto the conjugate pad and the sample flows by capillarity throughout the strip until reaching the test line, where a sandwich-like immunocomplex takes place due to the presence of antibody-functionalized QDs (Ab-QDs) onto the test line. Eventually, GO is added as a revealing agent and the photoluminescence of those sites protected by the complex Ab-SiO2/Antigen/Ab-QDs will not be quenched, whereas those photoluminescent sites directly exposed are expected to be quenched by GO, including the control line, made of bare QDs, reporting that the assay occurred successfully. Hence, the photoluminescence of the test line is modulated by the formation of sandwich-like immunocomplexes. The proposed device achieves a limit of detection (LOD) of 1.35 ng mL−1 in standard buffer, which is lower when compared with conventional lateral flow technology reported by gold nanoparticles, including other amplification strategies. Moreover, the resulting device was proven useful in human serum analysis, achieving a LOD of 6.30 ng mL−1 in this complex matrix. This low-cost disposable and easy-to-use device will prove valuable for portable and automated diagnostics applications, and can be easily transferred to other analytes such as clinically relevant protein biomarkers.