Real-time monitoring of voltage shift based on enzymatically released pyrophosphate using phenylboronic acid-immobilized gate field-effect transistor

• Published in: Japanese Journal of Applied Physics Vol. 53; no. 4S; pp. 4 - 1-04EL04-3
• Main Authors: Nishida, Hirokazu, Takahashi, Kiyofumi, Tabuse, Yuki, Kambara, Hideki, Sakata, Toshiya
• Format: Journal Article
• Language: English
• Published: The Japan Society of Applied Physics 01-04-2014
• Subjects: Conjugation; Deoxyribonucleic acid; DNA polymerase; Electric potential; Field effect transistors; Gates; Monitoring; Semiconductor devices; Voltage
• ISSN: 0021-4922; 1347-4065
• DOI: 10.7567/JJAP.53.04EL04

Pyrophosphate (PPi) is ubiquitous in living cells and is often produced by enzymatic reactions, e.g., DNA synthesis by DNA polymerase. We have developed a novel detection system for the voltage shift associated with the change in PPi concentration resulting from an enzymatic reaction using a phenylboronic acid (PBA)-coated gate field-effect transistor (FET), since PBA coating is effective for detecting ion accumulation associated with PPi production from enzymatic reactions. To detect enzymatic reactions more efficiently, we employed the enzyme-electrode conjugation method using specific peptide sequences, which are spontaneously tethered to a gold substrate. The combination of the enzyme-electrode conjugation method with the charge detection using the PBA-coated FET enables the effective detection of enzymatic reactions.