An electrode array for electrochemical immuno-sensing using the example of impedimetric tenascin C detection

Abstract

Electrochemical biosensors allow simple, fast and sensitive analyte detection for various analytical problems. Especially immunosensors are favourable due to specificity and affinity of antigen recognition by the associated antibody. We present a novel electrode array qualified for parallel analysis and increased sample throughput. The chip has nine independent sample chambers. Each chamber contains a circular gold working electrode with a diameter of 1.9 mm that is surrounded by a ring-shaped auxiliary electrode with a platinum surface. The corresponding silver/silver chloride reference electrodes are embedded in a sealing lid. The chip is open to the full range of electrochemical real-time detection methods. Among these techniques, impedance spectroscopy is an attractive tool to detect fast and label-free interfacial changes originating from the biorecognition event at the electrode surface. The capabilities of the novel electrode array are demonstrated using the example of tumour marker tenascin C detection. This glycoprotein of the extracellular matrix is expressed in cancerous tissues, especially in solid tumours such as glioma or breast carcinoma. Electrodes covered with specific antibodies were exposed to tenascin C containing samples. Non-occupied binding sites were identified using a secondary peroxidase-conjugated antibody that generated an insoluble precipitate on the electrode in a subsequent amplification procedure. The charge transfer resistance obtained from impedimetric analysis of ferri-/ferrocyanide conversion at the electrode served as analytic parameter. This assay detected 14 ng (48 fmol) tenascin C that is sufficient for clinical diagnostics. The electrode surface could be regenerated at least 20-fold without loss of its analytical performance.