Structural Manipulation of Microwell for Enhancing Analytical Performance of Enzyme Immunoassay

In this investigation, a novel design for a well-plate structure was created to optimize antigen-antibody reactions.The main objective during the development process was to enhance the internal structure of the well plate and increase the surface area.To improve efficiency, the newly designed well-plate was conical in shape and featured internal protrusions, or fins, which increased the surface area per unit volume by 1.45 times compared to standard plates.The performance of the newly developed well plate was assessed using a sandwich CLEIA system, which demonstrated a detection limit approximately 2.5 times better than that of commercial products.Additionally, the coefficient of variation (CV%) was superior to that of commercial products, with inter-assay CV(%) 11 and intra-assay CV(%) 9, compared with interassay CV(%) 15 and intra-assay CV(%) 10 for commercial products.Furthermore, the newly designed well plate demonstrated higher reaction efficiency, even with smaller sample volumes (25~50 L) compared to the 50~100 L typically required by commercial well plates.The incorporation of fine patterns increases the number of active sites available for interaction with the samples, thereby significantly enhancing the reaction sensitivity and overall performance.