A novel assay using synthetic protein scaffolds to detect soluble lectin-like Oxidised Low Density Lipoprotein Receptor-1 in vascular disease

Abstract

Methods: The bacterium E.coli was used to produce recombinant Adhirons that recognised human sLOX-1. Different ELISA-based assays were analysed for the ability of Adhirons to bind and detect sLOX-1 in biological buffers and human plasma. Results: Hybrid antibody-sLOX-1-Adhiron and homogenous AdhironsLOX-1-Adhiron assays could detect sLOX-1 levels down to picogram per millilitre sensitivity. Analysis of sLOX-1 levels using conventional antibody or Adhiron-based assays suggests that (1) sLOX-1 levels are elevated in chronic disease states, (2) Adhirons have high affinity and specificity for sLOX-1, and (3) Adhiron-based assays are a viable alternative for monitoring sLOX-1 biomarker levels. Conclusions: This study demonstrates that Adhirons can act as probes for monitoring sLOX-1 levels in vascular disease states. Further refinement and optimisation is needed for monitoring sLOX-1 levels in human blood samples and relating this to patient disease status. These studies provide a platform for using sLOX-1 as a biomarker in acute and chronic vascular disease.