Electrochemical Impedance-Based Detection of Pancreatic Cancer Biomarker Glypican1 and Mucin1 Using Electric Field-Lysed Extracellular Vesicles for Analysis: A Proof of Concept

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Abstract
Glypican1 and Mucin1 antigens are prominent biomarkers for the prognosis and diagnosis of pancreatic cancer. Their presence within the extracellular vesicles (EV) opens the possibilities for oncology care through the development of minimally invasive biomarker-assisted screening tools. Traditionally, EV antigen quantification relies on ultracentrifugation and chemical lysis, which are time-consuming, equipment-dependent, and often compromise EV integrity, damaging surface intact biomarkers. This study integrates EV isolation and electric field (EF) lysis into a unified platform. The lysates were then analyzed using an electrochemical impedance spectroscopy (EIS) -based sensor to detect GPC1 and MUC1. ELISA confirms the EF lysis of the immobilized EV and shows an increase in the antigen concentration by 2.5 times (compared to pre-lysed sample). Hence, EF lysis makes the sensor more sensitive than traditional methods. To enhance the electric lysis process, we applied varying voltages of a sinusoidal signal to the SPE-immobilized EVs. The lysate was subsequently used to quantify the GPC1 and MUC1 antigens through EIS. The results indicate that a 50 mV sinusoidal signal is sufficient to effectively lyse EVs, confirmed by western blotting. The NTA results showed the successful isolation of 109 EVs from 100 μL of serum using CD63 antibody. The developed EIS sensor can detect GPC1 and MUC1 with a LOD of 0.053 pg/mL and 0.033 pg/mL respectively from EV lysate, showing minimal non-specific binding in the negative control. Beyond GPC1 and MUC1, the approach is adaptable for detecting other EV-associated biomarkers, enabling broader applications in early cancer detection and disease monitoring.
Funding Information
  • Abdul Kalam Technology Innovation National Fellowship through the Science and Engineering Research Board, Department of Science and Technology (INAE/121/AKF/42)

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