Electrochemical nucleic acid hybridization biosensor based on poly(L-Aspartic acid)-modified electrode for the detection of short oligonucleotide sequences related to hepatitis C virus 1a

• Published in: Preparative biochemistry & biotechnology Vol. 49; no. 9; pp. 900 - 907
• Main Authors: Donmez, Soner, Çağdaş, Leman, Arslan, Halit, Arslan, Fatma
• Format: Journal Article
• Language: English
• Published: England Taylor & Francis 2019; Taylor & Francis Ltd
• Subjects: Acids; Aspartic acid; Avidin; Biomolecules; Biosensing Techniques - instrumentation; Biosensors; Biotin; DNA Probes - chemistry; DNA Probes - genetics; Electrochemical Techniques - instrumentation; Electrochemistry; Electrodes; Fabrication; HCV 1a; Hepacivirus - genetics; Hepacivirus - isolation & purification; Hepatitis; Hepatitis C; Hepatitis C - virology; Humans; Hybridization; Immobilization; Immobilized Nucleic Acids - chemistry; Immobilized Nucleic Acids - genetics; Morphology; Nucleic acid biosensor; Nucleic Acid Hybridization; Nucleic acids; Oligonucleotides; Oligonucleotides - analysis; Oligonucleotides - genetics; Peptides - chemistry; Poly (L-aspartic acid); Polymerase chain reaction; Polymerization; Scanning electron microscopy; Shelf life; Square waves; Viruses; HCV 1a; Poly (L-aspartic acid); Nucleic acid biosensor
• ISSN: 1082-6068; 1532-2297
• DOI: 10.1080/10826068.2019.1636279

This work describes, for the first time, the fabrication of poly(L-aspartic acid) (PAA) film modified pencil graphite electrode (PGE) for the detection of hepatitis C Virus 1a (HCV1a). The presence of PAA on the electrode surface can provide free carboxyl groups for covalent binding of biomolecules. The PGE surface was first coated with PAA via electropolymerization of the L-aspartic acid, and avidin was subsequently attached to the PAA modified electrode by covalent attachment. Biotinylated HCV1a probes were immobilized on avidin/PAA/PGE via avidin-biotin interaction. The morphology of PAA/PGE was examined using a scanning electron microscope. The hybridization events were monitored with square wave voltammetry using Meldola's blue (MDB). Compared to non-complementary oligonucleotide sequences, when hybridization was carried out between the probe and its synthetic targets or the synthetic polymerase chain reaction analog of HCV1a, the highest MDB signal was observed. The linear range of the biosensor was 12.5 to 100 nM and limit of detection was calculated as 8.7 nM. The biosensor exhibited favorable stability over relatively long-term storage. All these results suggest that PAA-modified electrode can be used to nucleic acid biosensor application and electropolymerization of L-aspartic acid can be considered as a good candidate for the immobilization of biomolecules.