The detection of cancer antigens is a major aim of cancer research in order to develop better patient management through early disease detection. Many cancers including prostate, lung, and ovarian secrete a protein disulfide isomerase protein named AGR2 that has been previously detected in urine and plasma using mass spectrometry. Here we determine whether a previously developed monoclonal antibody targeting AGR2 can be adapted from an indirect two-site ELISA format into a direct detector using solid-phase printed gold electrodes. The screen-printed gold electrode was surface functionalized with the anti-AGR2 specific monoclonal antibody. The interaction of the recombinant AGR2 protein and the anti-AGR2 monoclonal antibody functionalized electrode changed its electrochemical impedance spectra. Nyquist diagrams were obtained after incubation in an increasing concentration of purified AGR2 protein with a range of concentrations from 0.01 fg/mL to 10 fg/mL. In addition, detection of the AGR2 antigen can be achieved from cell lysates in medium or artificial buffer. These data highlight the utility of an AGR2-specific monoclonal antibody that can be functionalized onto a gold printed electrode for a one-step capture and quantitation of the target antigen. These platforms have the potential for supporting methodologies using more complex bodily fluids including plasma and urine for improved cancer diagnostics.

Cambridge Oesophagogastric Centre Cambridge University Hospitals NHS Foundation Trust Cambridge CB2 0QQ UK

Institute of Biotechnology and Molecular Medicine 3 Trzy Lipy St 80 172 Gdansk Poland

Institute of Genetics and Molecular Medicine University of Edinburgh Edinburgh EH4 2XR UK

International Centre for Cancer Vaccine Science University of Gdansk Kładki 24 St 80 822 Gdańsk Poland

Research Centre for Applied Molecular Oncology Masaryk Memorial Cancer Institute 65653 Brno Czech Republic

SensDx 14b Postępu St 02 676 Warszawa Poland

UKM Medical Centre UKM Medical Molecular Biology Institute Universiti Kebangsaan Malaysia Cheras Kuala Lumpur 56000 Malaysia

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Anterior gradient proteins in gastrointestinal cancers: from cell biology to pathophysiology