biostudies-literatureSingampalli KLWellcome Trust340246https://www.ebi.ac.uk/biostudies/studies/S-EPMC9458869biostudies-literatureUnknown1225Protein-based diagnostics are the standard of care for screening and diagnosing a broad range of diseases and medical conditions. The current gold standard method for quantifying proteins in clinical specimens is the enzyme-linked immunosorbent assay (ELISA), which offers high analytical sensitivity, can process many samples at once, and is widely available in many diagnostic laboratories worldwide. However, running an ELISA is cumbersome, requiring multiple liquid handling and washing steps, and time-intensive (∼2 - 4 h per test). Here, we demonstrate a unique magneto-ELISA that utilizes dually labeled magnetic nanoparticles (DMPs) coated with horseradish peroxidase (HRP) and an HRP-conjugated detection antibody, enabling rapid immunomagnetic enrichment and signal amplification. For proofAnalytica chimica actaRapid magneto-enzyme-linked immunosorbent assay for ultrasensitive protein detection.PMC9458869215826/Z/19/ZLi JSingampalli KLLillehoj PBfalseRapid magneto-enzyme-linked immunosorbent assay for ultrasensitive protein detection.Protein-based diagnostics are the standard of care for screening and diagnosing a broad range of diseases and medical conditions. The current gold standard method for quantifying proteins in clinical specimens is the enzyme-linked immunosorbent assay (ELISA), which offers high analytical sensitivity, can process many samples at once, and is widely available in many diagnostic laboratories worldwide. However, running an ELISA is cumbersome, requiring multiple liquid handling and washing steps, and time-intensive (∼2 - 4 h per test). Here, we demonstrate a unique magneto-ELISA that utilizes dually labeled magnetic nanoparticles (DMPs) coated with horseradish peroxidase (HRP) and an HRP-conjugated detection antibody, enabling rapid immunomagnetic enrichment and signal amplification. For proof2022-01-01T00:00:00Z2022 Sep2025-04-20T00:40:34.326Z2025-04-20T00:40:34.326ZS-EPMC94588693603824210.1016/j.aca.2022.340246