Abstract/Summary

The performance of biosensors is often optimised in buffers, which brings inconsistencies during applications with biological samples. Current strategies for minimising sample (matrix) interference are complex to automate and miniaturise, involving e.g. sample dilution or recovery of serum/plasma. This study shows the first systematic study using hundreds of actual microfluidic immunoassay fluoropolymer strips to understand matrix interference in microflow systems. As many interfering factors are assay-specific, we have explored matrix interference for a range of enzymatic immunoassays, including a direct mIgG/anti-mIgG, a sandwich cancer biomarker PSA and a sandwich inflammatory cytokine IL-1β. Serum matrix interference was significantly affected by capillary antibody surface coverage, suggesting for the first time the main cause of serum matrix effect is low-affinity serum components (e.g. auto-antibodies) competing with high-affinity antigen for the immobilised antibody. Additional experiments carried out with different capillary diameters confirmed the importance of antibody surface coverage in managing matrix interference. Building on these findings we propose a novel analytical approach where antibody surface coverage and sample incubation times are key for eliminating and/or minimising serum matrix interference, consisting in bioassay optimization carried out in serum instead of buffer, without compromising the performance of the bioassay nor adding extra cost nor steps. This will help establishing a new route towards faster development of modern point-of-care tests and effective biosensors development.