Enhanced ELISA Detection by Target Protein Enrichment with MIP: A New Strategy for Ultra-Sensitive Protein Detection

Background Ultra-sensitive detection of biomarkers is critical for disease diagnosis and other applications. Immunoassays, particularly enzyme-linked immunosorbent assay (ELISA), are commonly used for protein biomarker detection but often suffer from insufficient sensitivity at low concentrations. Enhancing the detection capacity of ELISA for early diagnosis is therefore essential. Results Herein, a simple and cost-effective strategy was developed to enhance ELISA performance by pre-enriching target proteins using molecularly imprinted polymers (MIPs). MIPs, synthesized with high absorption capacity and specificity, were prepared using TrF (Transferrin) as the template and polypeptide as crosslinker. Target protein was pre-enriched through large-volume adsorption followed by small-volume elution. This pre-enrichment process resulted in an 8.25-fold increase in protein concentration, significantly improving ELISA sensitivity and reducing the limit of detection (LOD) by nearly an order of magnitude. The enhanced detection performance was validated in terms of accuracy, precision, and linearity. Significance This method, which combines MIP-based protein enrichment with ELISA detection, offers a promising and cost-effective solution for enhancing ELISA performance. It holds great potential as a widely applicable strategy for the sensitive detection of biomarkers in disease diagnostics.