Gold-Loaded Nanoporous Iron Oxide Nanocubes: A Novel Dispersible Capture Agent for Tumor-Associated Autoantibodies Analysis in Serum

Abstract

Autoantibodies are produced against tumor associated antigens (TAAs) long before the appearance of any symptoms and thus can serve as promising, non-invasive biomarkers for early diagnosis of cancer. Current conventional methods for autoantibody detection are highly invasive and mostly provide diagnosis in the later stages of cancer. Herein we report a new electrochemical method for early detection of p53 autoantibodies against colon cancer using a strategy that combines the strength of gold-loaded nanoporous iron oxide nanocubes (Au@NPFe2O3NC)-based capture and purification while incorporating the inherent simplicity, inexpensive, and portable nature of the electrochemical and naked-eye colorimetric readouts. After the functionalisation of Au@NPFe2O3NC with p53 antigens, our method utilises a two-step strategy that involves (i) magnetic capture and isolation of autoantibodies using p53/ Au@NPFe2O3NC as ‘dispersible nanocapture agent’ in serum samples and (ii) subsequent detection of autoantibodies through peroxidase-catalyzed reaction on a commercially available disposable screen-printed electrode or naked-eye detection in an eppendorf tube. The method has demonstrated a good sensitivity (LOD= 0.02 U/mL) and reproducibility (relative standard deviation, % RSD = < 5%, n=3) for detecting p53 autoantibodies in serum and was also successfully applied to analyse a small cohort of clinical samples obtained from colorectal cancer. We believe that highly inexpensive, rapid, sensitive, and specific nature of our assay could potentially aid in the development of an early diagnostics tool for cancer and related diseases.