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Microchannel Immunoassays with Electrochemical Detection in Devices Constructed from Low Temperature Co-fired Ceramic and Screen Printed Gold

E.S.O. Fakunle and I. Fritsch
University of Arkansas, US

Keywords:
LTCC, microchannels, immunassays, electrochemistry

Abstract:
We report the first self-contained electrochemical enzyme linked immunosorbent assay in microchannel devices constructed from low temperature co-fired ceramic (LTCC) with screen-printed gold for fast and sensitive assays. No one has previously reported a device constructed from LTCC with integrated electrodes for combined immobilization and electrochemical detection. The LTCC microchannel devices used for the immunoassays contain four integrated individually addressable screen-printed gold electrodes. The total channel volume is under 0.8mL. The entire immunoassembly and electrochemical detection is carried out within the microchannel device.We demonstrate the microchannel immunoassays using mouse IgG as the model analyte. The enzyme linked immunosorbent assay was developed inside the microchannel using a protocol whereby self-assembled monolayers (SAMs) of mercatoundecanol were formed on the screen-printed gold on the microchannel wall and served as the interface for the covalent attachment of rat anti-mouse IgG. Mouse IgG was captured from the sample by the antibody, and then a secondary antibody (rat anti-mouse IgG) that is conjugated to alkaline phosphatase completed the sandwich-type immunoassembly. After addition of p-aminophenylphosphate, p-aminophenol is generated enzymatically by immobilized antibody conjugated alkaline phosphatase and is detected electrochemically using the microchannel electrodes.

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