Electrochemical detection of enzyme labeled DNA based on disposable pencil graphite electrode


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Kara P. , ERDEM A. , GIROUSI S., OZSOZ M.

JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS, cilt.38, ss.191-195, 2005 (SCI İndekslerine Giren Dergi) identifier identifier identifier

  • Cilt numarası: 38 Konu: 1
  • Basım Tarihi: 2005
  • Doi Numarası: 10.1016/j.jpba.2004.12.011
  • Dergi Adı: JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
  • Sayfa Sayıları: ss.191-195

Özet

Electrochemical biosensor for the detection of DNA hybridization using the reduction signal of a-naphthol is described. A pencil graphite electrode was used as a working electrode. Capture probes were covalently attached on to the pencil graphite electrode surface (PGE) at the 5' end amino group by using N-(dimethylamino)propyl-N'-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysulfosuccinimide (NHS) as a coupling agent on to PGE. After capture probe immobilization on to PGE surface; probe was hybridized with complementary biotinylated oligonucleotide. Alkaline phosphatase labeled with extravidin (Ex-AP) binds to biotinylated hybrid via biotin-avidin interaction. alpha-Naphthyl phosphate (alpha-NAP) was added and the reaction between alkaline phosphatase (AP) and alpha-NAP was occurred consequently as a substrate of AP, alpha-NAP reduction signal was obtained from this reaction, at -0.100 V by using differential pulse voltammetry (DPV). Other experimental parameters were studied such as; optimizations of hybridization time, and the concentrations of capture probe, biotinylated oligonucleotide and enzyme. (c) 2005 Elsevier B.V All rights reserved.