Single-step covalent immobilization of oligonucleotides onto solid surface

Abstract

In this communication, we report a simple, single-step method for covalent immobilization of oligonucleotide probes onto an activated polystyrene surface. Polystyrene surface is activated by a photolinker, 1-fluoro-2-nitro-4-azidobenzene (FNAB) in a photochemical reaction by UV light. Amine-modified oligonucleotide probe is covalently immobilized to the activated surface by displacing the labile fluoro group of the activated polystyrene surface by the primary amino group of the aminated oligonucleotide probe. Biotinylated complementary oligonucleotide target is hybridized with the immobilized probe. The hybridized target is detected by binding streptavidin-peroxidase followed by assaying the enzyme for colorimetric signal measurement. The covalently immobilized oligonucleotide probe shows high selectivity in subsequent hybridization processes with the complementary target and clearly discriminates with single-base mismatched oligonucleotide targets. The method can immobilize amine-or thiol-modified oligonucleotide onto FNAB-activated polymer surface. The method could be potentially useful for designing DNA based biosensors or biochips and also in microtiter plate based assay systems in a cost effective manner.

Gold nanoparticles assisted characterization of amine functionalized polystyrene multiwell plate and glass slide surfaces

“…Immobilization of biomolecules on the solid surface was the key step in the diagnosis of infectious and genetic diseases, analysis of gene expression, forensic science, toxicology, drug discovery, biosensing, and environmental monitoring (Kannoujia et al 2010;North et al 2010). In general, polystyrene (PS), polypropylene, polycarbonate, and glass slide (GS) have been used as a solid materials for the immobilization of biomolecules (DNA and protein) because of their versatile chemical, physical, and surface adherence properties.…”

Hydrogel Based Biosensors for In Vitro Diagnostics of Biochemicals, Proteins, and Genes

“…Conventionally, solid surfaces such as glass, silicon, gold, and plastics have been widely used for the immobilization of various oligonucleotides for biosensing applications . However, there is a fundamental limitation of 2D surfaces that lower the sensitivity of detection systems when probe DNA is confined to the surface of substrates.…”

“…It is likely that the efficiency and kinetics of hybridization are highly influenced by the surface density of DNA strands and their relative mobility . Unlike 2D surfaces, the hydrogel can provide an increased accessibility of gel‐immobilized nucleic acids for more rapid and efficient base‐pair hybridization . Here, we introduce various approaches to combining different sensor platforms with 3D hydrogels for sensitive detection of disease specific genes and nucleic acid (NA).…”

Analysis of global and sequential mass transfers for the adsorption of atrazine and simazine onto granular activated carbons from a hydrodynamic column

“…A close relationship between the surface chemical functions (SCFs) and GMT for the GAC prepared using the same starting material was justified by considering that different SCFs may arise due to kinetic effects and/or ageing, involving modifications of the surface groups. 30,31 Both the SCFs for TAS and GRW2 (the two GACs originating from coconut shell) are comparable, and have a relatively important SCFs contribution compared with the SCFs for F400 (the GAC originating from coal). 1 The nature of the SCFs is typified by the polar functional groups, so the influence of SCFs on the GMT is dependent on the polarity of the adsorbate molecule.…”